CRL-11609 RWPE-1 人正常前列腺上皮細(xì)胞;原代細(xì)胞|細(xì)胞系|細(xì)胞株|菌種;細(xì)胞庫管理規(guī)范,提供的細(xì)胞株背景清楚,提供參考文獻(xiàn)和*培養(yǎng)條件
CRL-11609 RWPE-1 人正常前列腺上皮細(xì)胞
Growth Properties: adherent
Organism: Homo Sapiens (human)
Propagation: Complete growth medium:K-SFM +0.05mg/ml BPE + 5ng/ml EGF
Temperature: 37.0℃
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
注:K-SFM (GIBCO 17005-042)含有BPE和EGF
Subculturing: Protocol: Remove medium, and rinse with PBS (pH7.2). Remove the solution and add 1 to 2 ml 0.05% trypsin-0.02% EDTA solution .Allow the flask to sit at 37.0℃ until the cells detach.Add fresh culture medium, aspirate and dispense into new culture flasks.
Subculturing ration: 1:3 to 1:5 is recommended.
Medium renewal: Every 2 days.
Preservation: Freeze medium: Complete growth medium + 40% NBS + 10% DMSO
Storage temperature: liquid nitrogen vapor temperature.
1. 接收到細(xì)胞,肉眼觀察細(xì)胞培養(yǎng)基顏色,顯微鏡觀察細(xì)胞生長(zhǎng)情況,并對(duì)細(xì)胞進(jìn)行不同倍數(shù)拍照(建議收到時(shí)的培養(yǎng)瓶拍一張照片,顯微鏡拍收到時(shí)的細(xì)胞100X,200X各一張)。
2. 用75%的酒精消毒(建議配置75%的酒精的水是滅菌過的)。
3. 嚴(yán)格無菌操作,打開細(xì)胞培養(yǎng)瓶。
4. 將細(xì)胞培養(yǎng)瓶?jī)?nèi)的培養(yǎng)基用吸管*吸出(嚴(yán)禁直接傾倒),放入另一無菌容器中(建議換新的培養(yǎng)基培養(yǎng)細(xì)胞)。
5. 用PBS 2-3ml/次輕輕洗細(xì)胞表面,洗3-4次遍,加入0.05%胰酶-EDTA1-1.5ml,顯微鏡下觀察細(xì)胞變圓,輕輕拍打培養(yǎng)瓶直到細(xì)胞脫落,加入終止液終止。
6. 1000rpm,5min離心,重懸細(xì)胞。
7. 換新的細(xì)胞培養(yǎng)瓶,37℃,5%CO2培養(yǎng)。