Hs683 HS683细胞 细胞株培养

 标题名称：Hs683 HS683细胞 细胞株培养

英文描述：

General informations

Culture conditions

Culture methods

Complete Growth Medium

The complete specialist medium for Hs683 cell line（Cat: ROCHEN PHARMA-CL-376) is ROCHEN PHARMA BIOSCIENCE uniquely formulated according to cell-growth recommendations of original cell line depositors . In some cases, media formulations differ slightly from other commercially available equivalents. Using ROCHEN PHARMA complete specialist medium is the best way to guarantee robust cell growth, ensuring that you'll have a supply of cells when you need them. The base medium for this cell line is ROCHEN PHARMA-formulated Dulbecco's Minimum Essential Medium (SH30243.01B). To make the complete growth medium, add the following components to the base medium: fetal bovine serum(ROCHEN PHARMA --formulated ) to a final concentration of 10%.

Subculturing

Volumes used in this protocol are for 25 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium.

2. Rinse the cell layer twice with 0.25% (w/v) Trypsin -0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.

3. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
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 37ºC to facilitate dispersal.

4. Add 4.0 to 6.0 mL 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°C.

Subc*tion Ratio

A subc*tion ratio of 1:2 is recommended.
Medium Renewal

Twice to3 times per week.

References

1. Owens RB, et al. Epithelial cell cultures from normal and cancerous human tissues. J. Natl. Cancer Inst. 56: 843-849, 1976. PubMed: 176412

2. Olopade OI, et al. Molecular analysis of deletions of the short arm of chromosome 9 in human gliomas. Cancer Res. 52: 2523-2529, 1992. PubMed: 1568221

3. Gershwin ME, et al. Immunobiology of heterotransplanted human tumors in nude mice. J. Natl. Cancer Inst. 58: 1455-1463, 1977. PubMed: 857033

4. Debinski W, et al. Receptor for interleukin (IL) 13 does not interact with IL4 but receptor for IL4 interacts with IL13 on human glioma cells. J. Biol. Chem. 271: 22428-22433, 1996. PubMed: 8798406

Hs683 HS683细胞 细胞株培养

收到 HS683细胞 处理方式：

新鲜细胞到货处理方式，客户收到细胞后按以下方式操作：

取出25cm2培养瓶，75%酒精消毒，拆下封口膜，放入37℃，5% CO2细胞培养箱中静置2-3h，以稳定细胞状态。

待细胞达到80%汇合时准备进行传代培养。

细胞传代，吸出25cm2培养瓶中的培养基,用PBS清洗细胞一次；添加0.25%胰蛋白酶消化液约1ml至培养瓶中，37℃温浴3min左右；倒置显微镜下观察，待细胞回缩变圆后吸弃消化液，再加入*培养液终止消化；用吸管轻轻吹打混匀，按1:2适当的比例进行接种传代（原代细胞必须以1:2方式传代），然后补充新鲜的*培养基至5ml，放入37℃，5% CO2细胞培养箱中培养；待细胞*贴壁后，培养观察。之后每隔2-3天更换新鲜的*培养基。

冻存管到货后处理方式，客户收到细胞产品后，请按照下述方法操作：

细胞冻存管公司多赠送一管，建议客户先复苏1支，如果复苏效果不理想，先与技术人员沟 通后再复苏另1支。

细胞收到后尽快复苏，有效期为一周。

收到冻存管后如不马上复苏，可以暂时放置于-80℃冰箱，不建议放入液氮罐。

复苏所用血清必须是胎牛血清。

复苏请按照细胞复苏 SOP 方式操作（细胞复苏 SOP 另行发送）。

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