最新中文字幕-精品国产99-国产精品福利在线观看-粉嫩av一区二区三区-日韩精品久久久久-美丽的姑娘国语视频-亚洲欧洲色-国产不卡毛片-三级黄毛片-免费福利在线-日韩av三区-欧美成年人网站-欧洲美女与动交ccoo-成人涩涩软件-秋霞影院午夜老牛影院-婷婷久久伊人-呦呦视频在线观看-www.天天综合-日韩精品高清视频-国产欧美综合在线-欧美日韩一区二区三区在线视频-a猛片免费播放-91九色蝌蚪国产-国产欧美日韩专区-嫩模被强到高潮呻吟不断

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which 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. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:5
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Wewer UM, et al. Heparan sulfate proteoglycans made by different basement-membrane- producing tumors have immunological and structural similarities. Differentiation 30: 61-67, 1985. PubMed: 2936642

Wewer U. Characterization of a rat yolk sac carcinoma cell line. Dev. Biol. 93: 416-421, 1982. PubMed: 7141105

Wewer U, et al. Laminin, a noncollagenous component of epithelial basement membranes synthesized by a rat yolk sac tumor. Cancer Res. 41: 1518-1524, 1981. PubMed: 7011537

. . Acta Pathol. Scand. Immunol. Sect. A, Immunol. 92: 275-283, 1984.

Oldberg A, et al. Isolation of a chondroitin sulfate proteoglycan from a rat yolk sac tumor and immunochemical demonstration of its cell surface localization. J. Biol. Chem. 256: 10847-10852, 1981. PubMed: 6793588

Lundstrom M, et al. Immunocytochemical and biochemical characterization of the Heymann nephritis antigenic complex in rat L2 yolk sac cells. Am. J. Pathol. 143: 1423-1435, 1993. PubMed: 8238258

Orlando RA, Farquhar MG. Identification of a cell line that expresses a cell surface and a soluble form of the gp330/receptor-associated protein (RAP) Heymann nephritis antigenic complex. Proc. Natl. Acad. Sci. USA 90: 4082-4086, 1993. PubMed: 8483924

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.