人妻少妇偷人精品无码丨色婷婷av久久久久久久丨欧美xxxx做受性欧美88丨欧洲女人牲交视频免费丨亚洲精品久久久av无码专区

(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. 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. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C without CO₂.

Brinkley BR, et al. Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro. Cancer Res. 40: 3118-3129, 1980. PubMed: 7000337

Cruciger Q, et al. Morphological, biochemical and chromosomal characterization of breast tumor lines from pleural effusions. In Vitro 12: 331, 1976.

Siciliano MJ, et al. Mutually exclusive genetic signatures of human breast tumor cell lines with a common chromosomal marker. Cancer Res. 39: 919-922, 1979. PubMed: 427779

Cailleau R, et al. Breast tumor cell lines from pleural effusions. J. Natl. Cancer Inst. 53: 661-674, 1974. PubMed: 4412247

Cailleau R, et al. Long-term human breast carcinoma cell lines of metastatic origin: preliminary characterization. In Vitro 14: 911-915, 1978. PubMed: 730202

Bates SE, et al. Expression of the transforming growth factor-alpha/epidermal growth factor receptor pathway in normal human breast epithelial cells. Endocrinology 126: 596-607, 1990. PubMed: 2294006

Dickstein B, et al. Increased epidermal growth factor receptor in an estrogen-responsive, adriamycin-resistant MCF-7 cell line. J. Cell. Physiol. 157: 110-118, 1993. PubMed: 8408230

Huguet EL, et al. Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54: 2615-2621, 1994. PubMed: 8168088

Satya-Prakash KL, et al. Cytogenetic analysis on eight human breast tumor cell lines: high frequencies of 1q, 11q and HeLa-like marker chromosomes. Cancer Genet. Cytogenet. 3: 61-73, 1981. PubMed: 7272986

Katayose Y, et al. Promoting apoptosis: a novel activity associated with the Cyclin-dependent kinase inhibitor p27. Cancer Res. 57: 5441-5445, 1997. PubMed: 9407946

Littlewood-Evans AJ, et al. The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. Cancer Res. 57: 5386-5390, 1997. PubMed: 9393764

Sheng S, et al. Maspin acts at the cell membrane to inhibit invasion and motility of mammary and prostatic cancer cells. Proc. Natl. Acad. Sci. USA 93: 11669-11674, 1996. PubMed: 8876194

De Vincenzo R, et al. Antiproliferative activity of colchicine analogues on MDR-positive and MDR-negative human cancer cell lines. Anticancer Drug Des. 13: 19-33, 1998. PubMed: 9474240

Soker S, et al. Characterization of novel vascular endothelial growth factor (VEGF) receptors on tumor cells that bind VEGF165 via its exon 7-endoded domain. J. Biol. Chem. 271: 5761-5767, 1996. PubMed: 8621443

Huguet EL, et al. Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54: 2615-2621, 1994. PubMed: 8168088