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

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

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.

Moore AE, et al. Culture characteristics of four permanent lines of human cancer cells. Cancer Res. 15: 598-602, 1955. PubMed: 13261081

Chen TR. Re-evaluation of HeLa, HeLa S3, and HEp-2 karyotypes. Cytogenet. Cell Genet. 48: 19-24, 1988. PubMed: 3180844

Toolan HW. Transplantable human neoplasms maintained in cortisone-treated laboratory animals: H.S. No. 1; H.Ep. No. 1; H.Ep. No. 2; H.Ep. No. 3; and H.Emb.Rh. No. 1. Cancer Res. 14: 660-666, 1954. PubMed: 13209540

Black FL, et al. Propagation of measles virus in a strain of human epidermoid cancer cells (Hep-2). Proc. Soc. Exp. Biol. Med. 93: 107-108, 1956. PubMed: 13370591

Tex. Rep. Biol. Med. 15: 588, 1957.

Moore AE. Tumorigenic activity of cultures. Ann. N.Y. Acad. Sci. 76: 497-505, 1958. PubMed: 13627875

St. Geme JW, et al. Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils. J. Bacteriol. 178: 6281-6287, 1996. PubMed: 8892830

Gromeier M, et al. Internal ribosomal entry site substitution eliminates neurovirulence in intergeneric poliovirus recombinants. Proc. Natl. Acad. Sci. USA 93: 2370-2375, 1996. PubMed: 8637880

Roller RJ, et al. Structure and function in the herpes simplex virus 1 RNA-binding protein US11: mapping of the domain required for ribosomal and nucleolar association and RNA binding in vitro. J. Virol. 70: 2842-2851, 1996. PubMed: 8627758

Herold BC, et al. Differences in the susceptibility of herpes simplex virus types 1 and 2 to modified heparin compounds suggest serotype differences in viral entry. J. Virol. 70: 3461-3469, 1996. PubMed: 8648678

Chang YE, et al. Properties of the protein encoded by the UL32 open reading frame of herpes simplex virus 1. J. Virol. 70: 3938-3946, 1996. PubMed: 8648731

Carter KL, et al. Characterization of the products of the UL43 gene of herpes simplex virus 1: potential implications for regulation of gene expression by antisense transcription. J. Virol. 70: 7663-7668, 1996. PubMed: 8892886

Carter KL, Roizman B. The promoter and transcriptional unit of a novel herpes simplex virus 1 alpha gene are contained in, and encode a protein in frame with, the open reading frame of the alpha22 gene. J. Virol. 70: 172-178, 1996. PubMed: 8523523

Jamaluddin M, et al. Inducible translational regulation of the NF-IL6 transcription factor by respiratory syncytial virus infection in pulmonary epithelial cells. J. Virol. 70: 1554-1563, 1996. PubMed: 8627674