Keay L
Biotechnol Bioeng. 1977 Mar;19(3):399-411. doi: 10.1002/bit.260190310.
The growth of L-60TM cells (a suspension culture adapted L-cell) on media composed of MEM (minimum essential medium (Eagle)) and bactopeptone autoclaved together or separately under a variety of conditions has veen determined. It has been found that MEM autoclaved with 0.5% bactopeptone at 15 psi for 20 min, cooled and then neutralized with NaHCO3, consistently supported good cell growth of L-60TM and L-929 cells. Similar results were obtained when the MEM and bactopeptone were autoclaved separately. The cells grew initially as a monolayer, subsequently becoming a stationary suspension. Some experiments were carried out with agitated suspension culture of L-60TM cells in the autoclaved MEM-bactopeptone combination with and without added methylcellulose and results were obtained which indicate that large scale suspension culture is possible in this system. Other peptones were also found to support cell growth. The autoclaved MEM-bactopeptone combination also supported the growth of Chang liver and Vero cells. The Chang liver cells rapidly dissociated from the plastic surface but the Vero cells remained sufficiently securely attached so that it was possible to grow them near to confluency in roller bottles.
已测定L - 60TM细胞(一种适应悬浮培养的L细胞)在由MEM(最低限度基本培养基(伊格尔))和细菌蛋白胨在各种条件下一起或分别高压灭菌的培养基上的生长情况。已发现,MEM与0.5%细菌蛋白胨在15磅力/平方英寸下高压灭菌20分钟,冷却后用碳酸氢钠中和,始终能支持L - 60TM和L - 929细胞良好生长。当MEM和细菌蛋白胨分别高压灭菌时也获得了类似结果。细胞最初以单层生长,随后变成静止悬浮液。对L - 60TM细胞在添加和不添加甲基纤维素的高压灭菌的MEM - 细菌蛋白胨组合中进行搅拌悬浮培养的一些实验,结果表明在该系统中大规模悬浮培养是可行的。还发现其他蛋白胨也能支持细胞生长。高压灭菌的MEM - 细菌蛋白胨组合也支持张氏肝细胞和非洲绿猴肾细胞的生长。张氏肝细胞迅速从塑料表面解离,但非洲绿猴肾细胞仍足够牢固地附着,以至于可以在滚瓶中使其生长至接近汇合状态。