Difference in growth factor requirements of rat 3Y1 cells among growth in mass culture,clonal growth in low density culture,and stimulation to enter S phase in resting culture |
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Authors: | Atsuyuki Okuda Yoshitsugu Kajiwara Genki Kimura |
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Institution: | (1) Department of Virology, Medical Institute of Bioregulation, Kyushi University 69, 812 Maidashi, Fukuoka, Japan;(2) Present address: Chemicals Inspection and Testing Institute, Japan, 19-14 Chuo-cho, 830 Kurume-shi, Japan |
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Abstract: | Summary A semiserum-free medium was developed for monolayer culture of rat 3Y1 fibroblastic cells. The main components of the developed
medium added to Dulbecco's modified Eagle's medium (DMEM) were insulin transferrin epidermal growth factor, poly-d-lysine, bovine albumin, oleic acid, and bovine α-globulin. In this medium, 3Y1 cells grew in mass culture at much the same
rate as in DMEM supplemented with 10% fetat bovine serum (FBS), and colonies, albeit of smaller sizes, diddform. Virally transformed
derivatives of 3Y1 (simian virus 40-3Y1, polyoma virus-3Y1 and adenovirus type 12-3Y1) also formed colonies in the semiserum-free
medium.
When trypsinized 3Y1 cells were seeded with the medium lacking α-globulin, neither growth in the mass culture nor clonal growth
in the low density culture (clonal growth) occurred. In this case, cell spreading was inhibited by albumin, and this inhibition
was overcome by adding α-globulin or treating dishes with serum. When albumin was excluded from the semiserum-free medium,
clonal growth did not occur, whereas growth in mass culture and stimulation of DNA synthesis in the resting mass culture (stimulation
of DNA synthesis) were not so drastically affected. When oleic acid was removed, growth in mass culture was inhibited considerably,
but no considerable effect was seen on clonal growth or on stimulation of DNA synthesis. In the absence of insulin, stimulation
of DNA synthesis was inhibited more markedly than when other components were removed, but such was not the case with growth
in mass culture and clonal growth. |
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Keywords: | semiserum-free medium growth factor requirement rat fibroblasts clonal growth stimulation of DNA synthesis |
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