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.

Several tests and experimental work have been done for identifying the best growth conditions and accumulated amount of lipid moiety in B. braunii, a microalga(UTEX 572) in terms of media composition. The specific growth rate was found to be the highest at 0.15 g/L-day when the phosphorus concentration was doubled with the other ingredients at the normal level. Experiments for varied media compositions revealed that the accumulation of lipid was the highest at 48% (dry cell weight based) in the nitrogen deficient medium and its corresponding specific growth rate was comparative to that in the normal BG 11 medium. In the bubble column experiments, carbon dioxide containing air produced four times more cell mass than air only. Light and glucose addition also enhanced cell mass with maximum, 1.8 g/L and accordingly 42% of lipid composition, which turned out to be a better strategy for higher lipid-producing microalgal culture.