5637(人膀胱癌細(xì)胞)
細(xì)胞名稱：5637 人膀胱癌細(xì)胞
組織來(lái)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?jī)?nèi)舊培養(yǎng)液，加少量PBS潤(rùn)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。控制吹打的力度，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?jī)?nèi)，加入新鮮培養(yǎng)基，置37℃溫箱培養(yǎng)，隔天觀察貼壁生長(zhǎng)情況。
二、懸浮細(xì)胞傳代
1.將細(xì)胞懸液轉(zhuǎn)移到無(wú)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?jī)?nèi)，加入新鮮培養(yǎng)基，置37℃溫箱培養(yǎng)。
