Caki-1人肾透明细胞癌皮肤转移细胞.docVIP

Caki-1人肾透明细胞癌皮肤转移细胞 Cat Number：KG346 For Research Use Only 一、组成: 组 份 KG346 细胞一瓶 25cm2 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、Complete growth medium (凯基货号：KGM 4892-500) 3、0.25% （W/V） Trypsin-0.53mM EDTA (凯基货号：KGY0012) 三、细胞简介: Organism: Homo sapiens (human) Tissue: kidney; Derived from metastatic site: skin Morphology: epithelial Propagation: Complete growth medium: McCOys5A +10%FBS+P/S Temperature: 37.0C Atmosphere: air, 95%; carbon dioxide (CO2), 5% Subculturing: Protocol: Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 ml 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°C to facilitate dispersal. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation ratio: A subcultivation ratio of 1:2 to 1:4 is recommendedMedium renewal: 2 to 3 times per week Preservation: Freeze medium: culture medium 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：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！