Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion

Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Jasmin Bagge [verfasserIn] Per Hölmich [verfasserIn] Freja Aabæk Hammer [verfasserIn] Jan O. Nehlin [verfasserIn] Kilian Vomstein [verfasserIn] Lars Blønd [verfasserIn] Lisbet Rosenkrantz Hölmich [verfasserIn] Kristoffer Weisskirchner Barfod [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2023

Schlagwörter:	Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion

Übergeordnetes Werk:	In: Journal of Experimental Orthopaedics - Wiley, 2015, 10(2023), 1, Seite n/a-n/a
Übergeordnetes Werk:	volume:10 ; year:2023 ; number:1 ; pages:n/a-n/a

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/s40634-023-00596-x

Katalog-ID:	DOAJ087544970

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520			\|a Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED.
650		4	\|a Stem cells
650		4	\|a Cryopreservation
650		4	\|a Biobanking
650		4	\|a Adipose tissue
650		4	\|a Microfragmentation
650		4	\|a Enzymatic digestion
653		0	\|a Orthopedic surgery
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700	0		\|a Freja Aabæk Hammer \|e verfasserin \|4 aut
700	0		\|a Jan O. Nehlin \|e verfasserin \|4 aut
700	0		\|a Kilian Vomstein \|e verfasserin \|4 aut
700	0		\|a Lars Blønd \|e verfasserin \|4 aut
700	0		\|a Lisbet Rosenkrantz Hölmich \|e verfasserin \|4 aut
700	0		\|a Kristoffer Weisskirchner Barfod \|e verfasserin \|4 aut
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matchkey_str	article:21971153:2023----::ucsflsltoovalseclsrmrorsremcorgetduaaioeisermainsihneseatrtscmaaietdoi
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allfields	10.1186/s40634-023-00596-x doi (DE-627)DOAJ087544970 (DE-599)DOAJb0dced9028e746918353d9038a41eb78 DE-627 ger DE-627 rakwb eng RD701-811 Jasmin Bagge verfasserin aut Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery Per Hölmich verfasserin aut Freja Aabæk Hammer verfasserin aut Jan O. Nehlin verfasserin aut Kilian Vomstein verfasserin aut Lars Blønd verfasserin aut Lisbet Rosenkrantz Hölmich verfasserin aut Kristoffer Weisskirchner Barfod verfasserin aut In Journal of Experimental Orthopaedics Wiley, 2015 10(2023), 1, Seite n/a-n/a (DE-627)792130855 (DE-600)2780021-0 21971153 nnns volume:10 year:2023 number:1 pages:n/a-n/a https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/article/b0dced9028e746918353d9038a41eb78 kostenfrei https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/toc/2197-1153 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2023 1 n/a-n/a
spelling	10.1186/s40634-023-00596-x doi (DE-627)DOAJ087544970 (DE-599)DOAJb0dced9028e746918353d9038a41eb78 DE-627 ger DE-627 rakwb eng RD701-811 Jasmin Bagge verfasserin aut Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery Per Hölmich verfasserin aut Freja Aabæk Hammer verfasserin aut Jan O. Nehlin verfasserin aut Kilian Vomstein verfasserin aut Lars Blønd verfasserin aut Lisbet Rosenkrantz Hölmich verfasserin aut Kristoffer Weisskirchner Barfod verfasserin aut In Journal of Experimental Orthopaedics Wiley, 2015 10(2023), 1, Seite n/a-n/a (DE-627)792130855 (DE-600)2780021-0 21971153 nnns volume:10 year:2023 number:1 pages:n/a-n/a https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/article/b0dced9028e746918353d9038a41eb78 kostenfrei https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/toc/2197-1153 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2023 1 n/a-n/a
allfields_unstemmed	10.1186/s40634-023-00596-x doi (DE-627)DOAJ087544970 (DE-599)DOAJb0dced9028e746918353d9038a41eb78 DE-627 ger DE-627 rakwb eng RD701-811 Jasmin Bagge verfasserin aut Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery Per Hölmich verfasserin aut Freja Aabæk Hammer verfasserin aut Jan O. Nehlin verfasserin aut Kilian Vomstein verfasserin aut Lars Blønd verfasserin aut Lisbet Rosenkrantz Hölmich verfasserin aut Kristoffer Weisskirchner Barfod verfasserin aut In Journal of Experimental Orthopaedics Wiley, 2015 10(2023), 1, Seite n/a-n/a (DE-627)792130855 (DE-600)2780021-0 21971153 nnns volume:10 year:2023 number:1 pages:n/a-n/a https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/article/b0dced9028e746918353d9038a41eb78 kostenfrei https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/toc/2197-1153 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2023 1 n/a-n/a
allfieldsGer	10.1186/s40634-023-00596-x doi (DE-627)DOAJ087544970 (DE-599)DOAJb0dced9028e746918353d9038a41eb78 DE-627 ger DE-627 rakwb eng RD701-811 Jasmin Bagge verfasserin aut Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery Per Hölmich verfasserin aut Freja Aabæk Hammer verfasserin aut Jan O. Nehlin verfasserin aut Kilian Vomstein verfasserin aut Lars Blønd verfasserin aut Lisbet Rosenkrantz Hölmich verfasserin aut Kristoffer Weisskirchner Barfod verfasserin aut In Journal of Experimental Orthopaedics Wiley, 2015 10(2023), 1, Seite n/a-n/a (DE-627)792130855 (DE-600)2780021-0 21971153 nnns volume:10 year:2023 number:1 pages:n/a-n/a https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/article/b0dced9028e746918353d9038a41eb78 kostenfrei https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/toc/2197-1153 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2023 1 n/a-n/a
allfieldsSound	10.1186/s40634-023-00596-x doi (DE-627)DOAJ087544970 (DE-599)DOAJb0dced9028e746918353d9038a41eb78 DE-627 ger DE-627 rakwb eng RD701-811 Jasmin Bagge verfasserin aut Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED. Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery Per Hölmich verfasserin aut Freja Aabæk Hammer verfasserin aut Jan O. Nehlin verfasserin aut Kilian Vomstein verfasserin aut Lars Blønd verfasserin aut Lisbet Rosenkrantz Hölmich verfasserin aut Kristoffer Weisskirchner Barfod verfasserin aut In Journal of Experimental Orthopaedics Wiley, 2015 10(2023), 1, Seite n/a-n/a (DE-627)792130855 (DE-600)2780021-0 21971153 nnns volume:10 year:2023 number:1 pages:n/a-n/a https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/article/b0dced9028e746918353d9038a41eb78 kostenfrei https://doi.org/10.1186/s40634-023-00596-x kostenfrei https://doaj.org/toc/2197-1153 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_636 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 10 2023 1 n/a-n/a
language	English
source	In Journal of Experimental Orthopaedics 10(2023), 1, Seite n/a-n/a volume:10 year:2023 number:1 pages:n/a-n/a
sourceStr	In Journal of Experimental Orthopaedics 10(2023), 1, Seite n/a-n/a volume:10 year:2023 number:1 pages:n/a-n/a
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topic_facet	Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion Orthopedic surgery
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authorswithroles_txt_mv	Jasmin Bagge @@aut@@ Per Hölmich @@aut@@ Freja Aabæk Hammer @@aut@@ Jan O. Nehlin @@aut@@ Kilian Vomstein @@aut@@ Lars Blønd @@aut@@ Lisbet Rosenkrantz Hölmich @@aut@@ Kristoffer Weisskirchner Barfod @@aut@@
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Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. 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callnumber-first	R - Medicine
author	Jasmin Bagge
spellingShingle	Jasmin Bagge misc RD701-811 misc Stem cells misc Cryopreservation misc Biobanking misc Adipose tissue misc Microfragmentation misc Enzymatic digestion misc Orthopedic surgery Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
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topic_title	RD701-811 Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion Stem cells Cryopreservation Biobanking Adipose tissue Microfragmentation Enzymatic digestion
topic	misc RD701-811 misc Stem cells misc Cryopreservation misc Biobanking misc Adipose tissue misc Microfragmentation misc Enzymatic digestion misc Orthopedic surgery
topic_unstemmed	misc RD701-811 misc Stem cells misc Cryopreservation misc Biobanking misc Adipose tissue misc Microfragmentation misc Enzymatic digestion misc Orthopedic surgery
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title	Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
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title_full	Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
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author_browse	Jasmin Bagge Per Hölmich Freja Aabæk Hammer Jan O. Nehlin Kilian Vomstein Lars Blønd Lisbet Rosenkrantz Hölmich Kristoffer Weisskirchner Barfod
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title_sort	successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
callnumber	RD701-811
title_auth	Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
abstract	Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED.
abstractGer	Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED.
abstract_unstemmed	Abstract Purpose To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis. Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED.
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container_issue	1
title_short	Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion
url	https://doi.org/10.1186/s40634-023-00596-x https://doaj.org/article/b0dced9028e746918353d9038a41eb78 https://doaj.org/toc/2197-1153
remote_bool	true
author2	Per Hölmich Freja Aabæk Hammer Jan O. Nehlin Kilian Vomstein Lars Blønd Lisbet Rosenkrantz Hölmich Kristoffer Weisskirchner Barfod
author2Str	Per Hölmich Freja Aabæk Hammer Jan O. Nehlin Kilian Vomstein Lars Blønd Lisbet Rosenkrantz Hölmich Kristoffer Weisskirchner Barfod
ppnlink	792130855
callnumber-subject	RD - Surgery
mediatype_str_mv	c
isOA_txt	true
hochschulschrift_bool	false
doi_str	10.1186/s40634-023-00596-x
callnumber-a	RD701-811
up_date	2024-07-04T02:07:38.592Z
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Methods Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at ‐80 °C in cryoprotectant‐medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence‐associated β‐galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin‐Red‐S and Oil‐Red‐O staining, respectively. Results Microfragmented AT from 7 patients was cryopreserved for a period of 46–150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p < 0.05). Low levels of senescence‐associated β‐galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31−CD34+CD45−CD90+CD146−), pericytes (CD31−CD34−CD45−CD90+CD146+), transitional pericytes (CD31−CD34+CD45−CD90+CD146+), and CD271+ stem cells (CD31−CD45−CD90+CD271+) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04). Conclusions Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. 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Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis – a comparative study of isolation by tissue explant culture and enzymatic digestion

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