5637(人膀胱癌細(xì)胞)

細(xì)胞名稱：5637  人膀胱癌細(xì)胞

組織來源：膀胱；癌

培養(yǎng)條件：RPMI-1640 +10% FBS；37℃，5% CO2

形      態(tài)：貼壁；上皮細(xì)胞樣

5637(人膀胱癌細(xì)胞)背      景：據(jù)報(bào)道，該細(xì)胞能生成 SCF、 IL-1、IL-3、IL-6、G-CSF、GM-CSF等。

General Information:

Organism: Homo sapiens, human

Tissue: urinary bladder

Culture Properties: adherent

Morphology: epithelial

Culture Method:

Complete Growth Medium: RPMI-1640 + 10% FBS + Penicillin/Streptomycin

5637(人膀胱癌細(xì)胞)Subculturing:

Volumes are given for a 25cm2

flask. Increase or decrease the

amount of dissociation medium needed proportionally for culture

vessels of other sizes.

5637(人膀胱癌細(xì)胞)1. Remove and discard culture medium.

2. Briefly rinse the cell layer with 0.25%(w/v) Trypsin-EDTA

solution to remove all traces of serum that contains trypsin

inhibitor.

3. Add 1.0 to 2.0mL of Trypsin-EDTA solution to flask and

observe cells under an inverted microscope until cell layer is

dispersed.

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°C, 5% CO2.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended

Cryopreservation:

Freeze medium: Complete growth medium supplemented with

10% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

5637(人膀胱癌細(xì)胞)傳代操作步驟

一、貼壁細(xì)胞傳代

1.提前將培養(yǎng)基、PBS放入37℃水浴鍋內(nèi)預(yù)熱，用75%酒精擦拭后再放入超凈臺(tái)內(nèi)。

2.吸除或倒掉細(xì)胞瓶內(nèi)舊培養(yǎng)液，加少量PBS潤洗細(xì)胞。

3.加入適量胰酶，使胰酶的量能蓋住細(xì)胞，37℃孵育，每隔2~3min顯微鏡下觀察，待貼壁細(xì)胞間間隙變大、細(xì)胞趨于圓形但還未漂起時(shí)棄去胰酶，加入新鮮培養(yǎng)基，晃動(dòng)細(xì)胞瓶，終止胰酶作用。

4.用吸管小心吹打貼壁的細(xì)胞，制成細(xì)胞懸液?？刂拼荡虻牧Χ?，避免產(chǎn)生大量的氣泡。

5.將細(xì)胞懸液分別接種到另外的2~3個(gè)細(xì)胞瓶內(nèi)，加入新鮮培養(yǎng)基，置37℃溫箱培養(yǎng)，隔天觀察貼壁生長情況。

二、懸浮細(xì)胞傳代

1.將細(xì)胞懸液轉(zhuǎn)移到無菌離心管內(nèi)，1000rpm離心5min。

2.棄去上清，加入新鮮的培養(yǎng)基，用吸管小心吹散沉淀，制成細(xì)胞懸液。

3.將細(xì)胞懸液分別接種到另外的2~3個(gè)細(xì)胞瓶內(nèi)，加入新鮮培養(yǎng)基，置37℃溫箱培養(yǎng)。