標(biāo)題名稱：Hs683 HS683細(xì)胞 細(xì)胞株培養(yǎng)

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細(xì)胞 細(xì)胞株培養(yǎng)

收到 HS683細(xì)胞 處理方式：

新鮮細(xì)胞到貨處理方式，客戶收到細(xì)胞后按以下方式操作：

取出25cm2培養(yǎng)瓶，75%酒精消毒，拆下封口膜，放入37℃，5% CO2細(xì)胞培養(yǎng)箱中靜置2-3h，以穩(wěn)定細(xì)胞狀態(tài)。

待細(xì)胞達(dá)到80%匯合時(shí)準(zhǔn)備進(jìn)行傳代培養(yǎng)。

細(xì)胞傳代，吸出25cm2培養(yǎng)瓶中的培養(yǎng)基,用PBS清洗細(xì)胞一次；添加0.25%胰蛋白酶消化液約1ml至培養(yǎng)瓶中，37℃溫浴3min左右；倒置顯微鏡下觀察，待細(xì)胞回縮變圓后吸棄消化液，再加入*培養(yǎng)液終止消化；用吸管輕輕吹打混勻，按1:2適當(dāng)?shù)谋壤M(jìn)行接種傳代（原代細(xì)胞必須以1:2方式傳代），然后補(bǔ)充新鮮的*培養(yǎng)基至5ml，放入37℃，5% CO2細(xì)胞培養(yǎng)箱中培養(yǎng)；待細(xì)胞*貼壁后，培養(yǎng)觀察。之后每隔2-3天更換新鮮的*培養(yǎng)基。

凍存管到貨后處理方式，客戶收到細(xì)胞產(chǎn)品后，請(qǐng)按照下述方法操作：

細(xì)胞凍存管公司多贈(zèng)送一管，建議客戶先復(fù)蘇1支，如果復(fù)蘇效果不理想，先與技術(shù)人員溝 通后再復(fù)蘇另1支。

細(xì)胞收到后盡快復(fù)蘇，有效期為一周。

收到凍存管后如不馬上復(fù)蘇，可以暫時(shí)放置于-80℃冰箱，不建議放入液氮罐。

復(fù)蘇所用血清必須是胎牛血清。

復(fù)蘇請(qǐng)按照細(xì)胞復(fù)蘇 SOP 方式操作（細(xì)胞復(fù)蘇 SOP 另行發(fā)送）。

熱賣：DC2.4細(xì)胞，SW48細(xì)胞，RS4:11細(xì)胞，KGN細(xì)胞，A2780細(xì)胞，A2780/DDP細(xì)胞，HL-1細(xì)胞，COV434細(xì)胞等

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