99精品久久99久久久久,亚洲国产成人精品无码一区二区,国产精品亚洲综合色区韩国,久久精品第一国产久精国产宅男66,亚洲国产日韩欧美一区二区三区,久久久久久精品免费无码无,国产中年熟女高潮大集合,亚洲AV无码成H在线观看

The cells are positive for keratin by immunoperoxidase staining.

(Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)

1. Culture never becomes 100% confluent. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 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 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. .
6. Incubate cultures at 37°C.

Chen TR, et al. Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins. Cancer Genet. Cytogenet. 10: 351-362, 1983. PubMed: 6652615

Chen TR, et al. Karyotype consistency in human colorectal carcinoma cell lines established in vitro. Cancer Genet. Cytogenet. 6: 93-117, 1982. PubMed: 7104989

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Leibovitz A, et al. Classification of human colorectal adenocarcinoma cell lines. Cancer Res. 36: 4562-4569, 1976. PubMed: 1000501

Santoro IM, Groden J. Alternative splicing of the APC gene and its association with terminal differentiation. Cancer Res. 57: 488-494, 1997. PubMed: 9012479

Tsao H, et al. Novel mutations in the p16/CDKN2A binding region of the Cyclin-dependent Kinase-4 gene. Cancer Res. 58: 109-113, 1998. PubMed: 9426066