• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.131-136
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• 1994

In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced