CHO-K1細(xì)胞-懸浮培養(yǎng)細(xì)胞

    CHO-K1細(xì)胞-懸浮培養(yǎng)細(xì)胞是指將復(fù)蘇CHO-K1細(xì)胞轉(zhuǎn)入T25 cm2細(xì)胞培養(yǎng)瓶，以F12K基礎(chǔ)培養(yǎng)基添加10%血清培養(yǎng)，待細(xì)胞長(zhǎng)滿(mǎn)單層后，加入 0.25 % 胰蛋白酶消化，傳代至裝有完全培養(yǎng)基的T25 cm2細(xì)胞培養(yǎng)瓶（Corning）中繼續(xù)培養(yǎng)，當(dāng)細(xì)胞鋪滿(mǎn)瓶底時(shí)，即得貼壁 CHO-K1 細(xì)胞。取貼壁CHO-K1 細(xì)胞，換液，加入15 mL含血清的完全基和 15 mL無(wú)血清CHO-K1專(zhuān)用培養(yǎng)基，2天后吸出一半培養(yǎng)液，加入等量的無(wú)血清CHO-K1，每2天重復(fù)一次此操作，直到胎牛血清濃度低于 1 %后，以基礎(chǔ)培養(yǎng)基B001 繼續(xù)培養(yǎng)，即培養(yǎng)基中血清濃度依次以 5 %，2 .5 %，1.25%，0.625 %，0 %降低。細(xì)胞在無(wú)血清CHO-K1專(zhuān)用培養(yǎng)基中密度達(dá)到5 ×106cells/ mL時(shí)，即得懸浮 CHO-K1 細(xì)胞。培養(yǎng)條件：傳代培養(yǎng)條件：37±1℃，120rmp，6.0%CO2

細(xì)胞名稱(chēng)：倉(cāng)鼠卵巢細(xì)胞（懸?。?

細(xì)胞簡(jiǎn)稱(chēng)：CHO-K1細(xì)胞-懸浮培養(yǎng)細(xì)胞

產(chǎn)品貨號(hào)：TCO-C047

種屬來(lái)源：倉(cāng)鼠

組織來(lái)源：卵巢

細(xì)胞形態(tài)：圓形

生長(zhǎng)特性： 懸浮生長(zhǎng)

培養(yǎng)體系：CD CHO Medium+Anti-Clumping Agent+L-Glutamine Solution

配套培養(yǎng)基貨號(hào)：TCO-G047

傳代比例：傳代密度：0.2~0.5×106cells/mL；每2-4天傳代一次

傳代周期：48-72 h

培養(yǎng)條件：氣相：94%空氣+6%CO2，溫度：37℃

凍存條件：60%基礎(chǔ)培養(yǎng)基+30%FBS+10%DMSO，液氮儲(chǔ)存

質(zhì)量檢測(cè)：細(xì)菌、真菌、支原體檢測(cè)均為陰性

參考文獻(xiàn)：

Puck TT, et al. Genetics of somatic mammalian cells III. Long-term cultivation of euploid cells from human and animal subjects. J. Exp. Med. 108: 945-956, 1958. PubMed: 13598821

Ham RG. Clonal growth of mammalian cells in a chemically defined, synthetic medium. Proc. Natl. Acad. Sci. USA 53: 288-293, 1965. PubMed: 14294058

Kao FT, Puck TT. Genetics of somatic mammalian cells, VII. Induction and isolation of nutritional mutants in Chinese hamster cells. Proc. Natl. Acad. Sci. USA 60: 1275-1281, 1968. PubMed: 5244736

Kao FT, Puck TT. Genetics of somatic mammalian cells. IV. Properties of Chinese hamster cell mutants with respect to the requirement for proline. Genetics 55: 513-524, 1967. PubMed: 6068403

Kao FT, et al. Complementation analysis on virus-fused Chinese hamster cells with nutritional markers. Science 164: 312-314, 1969. PubMed: 5813351

Kao F, et al. Genetics of somatic mammalian cells. X. Complementation analysis of glycine-requiring mutants. Proc. Natl. Acad. Sci. USA 64: 1284-1291, 1969. PubMed: 5271752

Mayr GA, Freimuth P. A single locus on human chromosome 21 directs the expression of a receptor for adenovirus type 2 in mouse A9 cells. J. Virol. 71: 412-418, 1997. PubMed: 8985365