PDF《PLC/PRF/5 人肝癌亚历山大细胞》

PLC/PRF/5 人肝癌亚历山大细胞 说明书修订日期:20181017 Cat Number:KG068 For Research Use Only

一、组成 组份KG068 细胞一瓶 25cm

2 细胞说明书

1 份 细胞培养注意事项

1 份

二、客户自备试剂

1、PBS (凯基货号:KGB5001)

2、Inomplete growth medium (凯基货号: KGM41500-500)

3、0.

25%(W/V)Trypsin-0.53mM EDTA (凯基货号:KGY0012)

4、FBS

三、细胞简介 Growth Properties: adherent Organism: Mouse Propagation: Complete growth medium: MEM + 10%FBS+ P/S Temperature: 37.0℃ Atmosphere: air, 95%;

carbon dioxide (CO2), 5% Subculturing: Protocol: Volumes are given for a

75 cm

2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning? T-75 flasks (catalog #430641) are recommended for subculturing this product. 1. Remove and discard culture medium. 2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. 3. Add 2.0 to 3.0ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37℃ to facilitate dispersal. 4. Add 6.0 to 8.0ml of complete growth medium and aspirate cells by gently pipetting. 5. Add appropriate aliquots of the cell suspension to new culture vessels. 6. Incubate cultures at 37℃. Subcultivation ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium renewal:

2 to

3 times per week Preservation: Freeze medium:FBS 90%;

DMSO, 10% Storage temperature: liquid nitrogen vapor phase

四、常见问题及解决方案

1、培养瓶有破裂,培养液有漏液:细胞极大可能会污染,所以我们会及时安排帮老师解决.

2、细胞漂浮(针对贴壁细胞) :培养瓶不开封,瓶口酒精擦拭后平躺放置在培养箱.次日观察,如细胞大部分又贴回 瓶底,表明细胞活力正常,剩余漂浮的细胞可以离心去掉,留10ml 培养液培养观察,细胞生长至汇合度 80%,进行 消化传代;

如细胞还是不贴壁,将细胞离心收集转到新培养瓶,原培养瓶加部分培养液继续培养,中间注意观察,我 们的技术人员会一直跟踪指导,直到问题解决. 客户收到细胞后请务必仔细阅读细胞注意事项,确保细胞的培养条件一致,如果由于培养条件不一 致导致细胞出现问题,责任由客户自行承担.由于运输的情况,所以极个别细胞会出现不稳定,客户收 到细胞后务必第一时间和我们联系,告知细胞具体情况,以便我们技术人员能及时有效的和老师沟通, 不胜感谢!