Document Detail


Characteristics and EGFP expression of goat mammary gland epithelial cells.
MedLine Citation:
PMID:  20113446     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
The aims of this study were (i) to establish a goat mammary gland epithelial (GMGE) cell line, and (ii) to determine if these GMGE cells could be maintained long-term in culture by continuous subculturing following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Primary culture of GMGE cells was achieved by outgrowth of migrating cells from the fragments of the mammary gland tissue of a lactating goat. The passage 16 GMGE cells were transfected with EGFP gene using lipofection. The expression of Cell keratins of epithelial cells in GMGE cells was test by immunofluorescence. Βeta-Casein gene mRNA was test for GMGE cells by RT-PCR. The results showed that when grown at low density on a plastic substratum, the GMGE cells formed islands, and when grown to confluency, the cells formed a monolayer and aggregated with the characteristic cobble-stone morphology of epithelial cells. GMGE cells could form dome-like structure which looked like nipple, and the lumen-like structures formed among the cells. Several blister-like structures appeared in the appearance of the cells. The GMGE cells contained different cell types, majority of the cells were short shuttle-like or polygon which were beehive-like. A part of cells were round and flat, a small number of cells were elongated. Some of the GMGE cells contained milk drops. The cell nuclei were round which had 2-4 obvious cores. The expression of Cell keratins demonstrated the property of epithelial cells in GMGE cells by immunofluorescence. The GMGE cells could express transcript encoding a Βeta-Casein protein. EGFP gene was successfully transferred into the GMGE cells, and the transfected cells could be maintained long-term in culture by continuous subculturing. In conclusion, we have established a EGFP gene transfected GMGE (ET-GMGE) cell line and maintained it long-term in culture by continuous subculturing.
Authors:
Y-M Zheng; X-Y He; Y Zhang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Reproduction in domestic animals = Zuchthygiene     Volume:  45     ISSN:  1439-0531     ISO Abbreviation:  Reprod. Domest. Anim.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-11-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9015668     Medline TA:  Reprod Domest Anim     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  e323-31     Citation Subset:  IM    
Copyright Information:
© 2010 Blackwell Verlag GmbH.
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
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