5637(人膀胱癌細胞)
細胞名稱：5637 人膀胱癌細胞
組織來源：膀胱；癌
培養條件：RPMI-1640 +10% FBS；37℃，5% CO2
形 態：貼壁；上皮細胞樣
5637(人膀胱癌細胞)背 景：據報道，該細胞能生成 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(人膀胱癌細胞)傳代操作步驟
一、貼壁細胞傳代
1.提前將培養基、PBS放入37℃水浴鍋內預熱，用75%酒精擦拭后再放入超凈臺內。
2.吸除或倒掉細胞瓶內舊培養液，加少量PBS潤洗細胞。
3.加入適量胰酶，使胰酶的量能蓋住細胞，37℃孵育，每隔2~3min顯微鏡下觀察，待貼壁細胞間間隙變大、細胞趨于圓形但還未漂起時棄去胰酶，加入新鮮培養基，晃動細胞瓶，終止胰酶作用。
4.用吸管小心吹打貼壁的細胞，制成細胞懸液。控制吹打的力度，避免產生大量的氣泡。
5.將細胞懸液分別接種到另外的2~3個細胞瓶內，加入新鮮培養基，置37℃溫箱培養，隔天觀察貼壁生長情況。
二、懸浮細胞傳代
1.將細胞懸液轉移到無菌離心管內，1000rpm離心5min。
2.棄去上清，加入新鮮的培養基，用吸管小心吹散沉淀，制成細胞懸液。
3.將細胞懸液分別接種到另外的2~3個細胞瓶內，加入新鮮培養基，置37℃溫箱培養。
