Data/hora: 06/06/2024 10:25:50

Provedor de dados: 	56	
País: 	Brazil	
Título: 	Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3	
Autores: 	Liu,Z.; Zhang,P.; Zhou,Y.; Qin,H.; Shen,T.	
Data: 	2010-05-01	
Ano: 	2010	
Palavras-chave: 	Human intestinal epithelial cell; Cell culture; Thermolysin; Endothelin-3; Cytokeratin; Brush border enzyme.	
Resumo: 	Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.	
Tipo: 	Info:eu-repo/semantics/article	
Idioma: 	Inglês	
Identificador: 	http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000500006	
Editor: 	Associação Brasileira de Divulgação Científica	
Relação: 	10.1590/S0100-879X2010007500036	
Formato: 	text/html	
Fonte: 	Brazilian Journal of Medical and Biological Research  v.43 n.5 2010	
Fonte: 	info:eu-repo/semantics/openAccess