A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species
Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for l...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Huifang Chen [verfasserIn] Juan Li [verfasserIn] Xiaoting Pan [verfasserIn] Zhichao Hu [verfasserIn] Jianfeng Cai [verfasserIn] Zijie Xia [verfasserIn] Nanshan Qi [verfasserIn] Shenquan Liao [verfasserIn] Zachary Spritzer [verfasserIn] Yinshan Bai [verfasserIn] Mingfei Sun [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2024 |
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| Übergeordnetes Werk: |
In: Parasites & Vectors - BMC, 2008, 17(2024), 1, Seite 17 |
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| Übergeordnetes Werk: |
volume:17 ; year:2024 ; number:1 ; pages:17 |
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| DOI / URN: |
10.1186/s13071-023-06090-8 |
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| Katalog-ID: |
DOAJ097433462 |
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| 245 | 1 | 2 | |a A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species |
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| 520 | |a Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract | ||
| 650 | 4 | |a Avian embryo | |
| 650 | 4 | |a Avian intestinal epithelial cell line (AIEC) | |
| 650 | 4 | |a E. tenella | |
| 650 | 4 | |a Culture model | |
| 653 | 0 | |a Infectious and parasitic diseases | |
| 700 | 0 | |a Juan Li |e verfasserin |4 aut | |
| 700 | 0 | |a Xiaoting Pan |e verfasserin |4 aut | |
| 700 | 0 | |a Zhichao Hu |e verfasserin |4 aut | |
| 700 | 0 | |a Jianfeng Cai |e verfasserin |4 aut | |
| 700 | 0 | |a Zijie Xia |e verfasserin |4 aut | |
| 700 | 0 | |a Nanshan Qi |e verfasserin |4 aut | |
| 700 | 0 | |a Shenquan Liao |e verfasserin |4 aut | |
| 700 | 0 | |a Zachary Spritzer |e verfasserin |4 aut | |
| 700 | 0 | |a Yinshan Bai |e verfasserin |4 aut | |
| 700 | 0 | |a Mingfei Sun |e verfasserin |4 aut | |
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10.1186/s13071-023-06090-8 doi (DE-627)DOAJ097433462 (DE-599)DOAJa364df52d3bd40fb803f3e0298c51520 DE-627 ger DE-627 rakwb eng RC109-216 Huifang Chen verfasserin aut A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract Avian embryo Avian intestinal epithelial cell line (AIEC) E. tenella Culture model Infectious and parasitic diseases Juan Li verfasserin aut Xiaoting Pan verfasserin aut Zhichao Hu verfasserin aut Jianfeng Cai verfasserin aut Zijie Xia verfasserin aut Nanshan Qi verfasserin aut Shenquan Liao verfasserin aut Zachary Spritzer verfasserin aut Yinshan Bai verfasserin aut Mingfei Sun verfasserin aut In Parasites & Vectors BMC, 2008 17(2024), 1, Seite 17 (DE-627)558690076 (DE-600)2409480-8 17563305 nnns volume:17 year:2024 number:1 pages:17 https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/article/a364df52d3bd40fb803f3e0298c51520 kostenfrei https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/toc/1756-3305 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 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_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 17 2024 1 17 |
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10.1186/s13071-023-06090-8 doi (DE-627)DOAJ097433462 (DE-599)DOAJa364df52d3bd40fb803f3e0298c51520 DE-627 ger DE-627 rakwb eng RC109-216 Huifang Chen verfasserin aut A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract Avian embryo Avian intestinal epithelial cell line (AIEC) E. tenella Culture model Infectious and parasitic diseases Juan Li verfasserin aut Xiaoting Pan verfasserin aut Zhichao Hu verfasserin aut Jianfeng Cai verfasserin aut Zijie Xia verfasserin aut Nanshan Qi verfasserin aut Shenquan Liao verfasserin aut Zachary Spritzer verfasserin aut Yinshan Bai verfasserin aut Mingfei Sun verfasserin aut In Parasites & Vectors BMC, 2008 17(2024), 1, Seite 17 (DE-627)558690076 (DE-600)2409480-8 17563305 nnns volume:17 year:2024 number:1 pages:17 https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/article/a364df52d3bd40fb803f3e0298c51520 kostenfrei https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/toc/1756-3305 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 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_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 17 2024 1 17 |
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10.1186/s13071-023-06090-8 doi (DE-627)DOAJ097433462 (DE-599)DOAJa364df52d3bd40fb803f3e0298c51520 DE-627 ger DE-627 rakwb eng RC109-216 Huifang Chen verfasserin aut A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract Avian embryo Avian intestinal epithelial cell line (AIEC) E. tenella Culture model Infectious and parasitic diseases Juan Li verfasserin aut Xiaoting Pan verfasserin aut Zhichao Hu verfasserin aut Jianfeng Cai verfasserin aut Zijie Xia verfasserin aut Nanshan Qi verfasserin aut Shenquan Liao verfasserin aut Zachary Spritzer verfasserin aut Yinshan Bai verfasserin aut Mingfei Sun verfasserin aut In Parasites & Vectors BMC, 2008 17(2024), 1, Seite 17 (DE-627)558690076 (DE-600)2409480-8 17563305 nnns volume:17 year:2024 number:1 pages:17 https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/article/a364df52d3bd40fb803f3e0298c51520 kostenfrei https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/toc/1756-3305 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 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_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 17 2024 1 17 |
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10.1186/s13071-023-06090-8 doi (DE-627)DOAJ097433462 (DE-599)DOAJa364df52d3bd40fb803f3e0298c51520 DE-627 ger DE-627 rakwb eng RC109-216 Huifang Chen verfasserin aut A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract Avian embryo Avian intestinal epithelial cell line (AIEC) E. tenella Culture model Infectious and parasitic diseases Juan Li verfasserin aut Xiaoting Pan verfasserin aut Zhichao Hu verfasserin aut Jianfeng Cai verfasserin aut Zijie Xia verfasserin aut Nanshan Qi verfasserin aut Shenquan Liao verfasserin aut Zachary Spritzer verfasserin aut Yinshan Bai verfasserin aut Mingfei Sun verfasserin aut In Parasites & Vectors BMC, 2008 17(2024), 1, Seite 17 (DE-627)558690076 (DE-600)2409480-8 17563305 nnns volume:17 year:2024 number:1 pages:17 https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/article/a364df52d3bd40fb803f3e0298c51520 kostenfrei https://doi.org/10.1186/s13071-023-06090-8 kostenfrei https://doaj.org/toc/1756-3305 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 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_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 17 2024 1 17 |
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novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for eimeria species |
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A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species |
| abstract |
Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract |
| abstractGer |
Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract |
| abstract_unstemmed |
Abstract Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines. Conclusions An AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract. Graphical Abstract |
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A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species |
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https://doi.org/10.1186/s13071-023-06090-8 https://doaj.org/article/a364df52d3bd40fb803f3e0298c51520 https://doaj.org/toc/1756-3305 |
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Juan Li Xiaoting Pan Zhichao Hu Jianfeng Cai Zijie Xia Nanshan Qi Shenquan Liao Zachary Spritzer Yinshan Bai Mingfei Sun |
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Juan Li Xiaoting Pan Zhichao Hu Jianfeng Cai Zijie Xia Nanshan Qi Shenquan Liao Zachary Spritzer Yinshan Bai Mingfei Sun |
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2024-07-04T01:14:47.097Z |
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