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)滿單層后,加入 0.25 % 消化,傳代至裝有培養(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含血清的基和 15 mL無(wú)血清CHO-K1專用培養(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:倉(cāng)鼠卵巢細(xì)胞(懸浮)
細(xì)胞簡(jiǎn)稱: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):
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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
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