title: Evaluation of embryonic age and the effects of different proteases on the isolation and primary culture of chicken intestinal epithelial cells in vitro.
abstract: The present study evaluates the effects of embryonic age and proteolytic enzymes on the isolation and primary culture of chicken enterocyte and to establish an effective technique for chicken intestinal epithelial cell (IEC) cultivation. Fourteen-day-old, 16-day-old and 18-day-old embryos (average weight: 52.23 ± 0.76 g, 50.86 ± 0.99 g, 48.98 ± 1.03 g) were the source for preparation of enterocyte culture, and trypsin-ethylene diamine tetraacetic acid, collagenase, thermolysin and combination of collagenase and thermolysin were used for digestion medium. Optimal culture protocols were determined by qualitative assays of proliferation. Cells isolated by using 14-day-old embryo and collagenase obtain the best attachment and growth in culture, and the production of continuously growing IEC cultures. Thus, we conclude that the use of collagenase as a dissociating enzyme and 14-day-old embryo as a source can be advantageously applied to the isolation of chicken IEC and this method may be useful for various applications and basic studies of the intestinal tract concerning such objects as physiology, immunology and toxicology.
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Yuan
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: He
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.
authors.forename: Jun-ming
authors.initials: JM
authors.lastname: Li
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.|Poultry Production Department, Faculty of Agriculture, Mansoura University, Mansoura, Egypt.
authors.forename: Mahmoud Mostafa
authors.initials: MM
authors.lastname: Azzam
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.
authors.forename: Jian-jun
authors.initials: JJ
authors.lastname: Lu
authors.affiliation: Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.
authors.forename: Xiao-ting
authors.initials: XT
authors.lastname: Zou
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D017364:Collagenases / Q000378:metabolism*; D003470:Culture Media; D004492:Edetic Acid / Q000494:pharmacology; D047108:Embryonic Development; D020895:Enterocytes / Q000166:cytology*; D007413:Intestinal Mucosa / Q000166:cytology* / Q000196:embryology* / Q000201:enzymology; D013820:Thermolysin / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology
chemical_list: D003470:Culture Media; D004492:Edetic Acid; D014357:Trypsin; D017364:Collagenases; D013820:Thermolysin
keywords: embryonic age; intestinal epithelial cell; isolation; primary culture; protease