5637(人膀胱癌细胞)
细胞名称：5637 人膀胱癌细胞
组织来源：膀胱；癌
培养条件：RPMI-1640 +10% FBS；37℃，5% CO2
形 态：贴壁；上皮细胞样
5637(人膀胱癌细胞)背 景：据报道，该细胞能生成 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(人膀胱癌细胞)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(人膀胱癌细胞)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(人膀胱癌细胞)传代操作步骤
一、贴壁细胞传代
1.提前将培养基、PBS放入37℃水浴锅内预热，用75%酒精擦拭后再放入超净台内。
2.吸除或倒掉细胞瓶内旧培养液，加少量PBS润洗细胞。
3.加入适量胰酶，使胰酶的量能盖住细胞，37℃孵育，每隔2~3min显微镜下观察，待贴壁细胞间间隙变大、细胞趋于圆形但还未漂起时弃去胰酶，加入新鲜培养基，晃动细胞瓶，终止胰酶作用。
4.用吸管小心吹打贴壁的细胞，制成细胞悬液。控制吹打的力度，避免产生大量的气泡。
5.将细胞悬液分别接种到另外的2~3个细胞瓶内，加入新鲜培养基，置37℃温箱培养，隔天观察贴壁生长情况。
二、悬浮细胞传代
1.将细胞悬液转移到无菌离心管内，1000rpm离心5min。
2.弃去上清，加入新鲜的培养基，用吸管小心吹散沉淀，制成细胞悬液。
3.将细胞悬液分别接种到另外的2~3个细胞瓶内，加入新鲜培养基，置37℃温箱培养。
