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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.

Fogh J. Human tumor cells in vitro. New York: Plenum Press; 1975.

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

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

Fogh J. Cultivation, characterization, and identification of human tumor cells with emphasis on kidney, testis, and bladder tumors. Natl. Cancer Inst. Monogr. 49: 5-9, 1978. PubMed: 571047

Heffelfinger SC, et al. SK HEP-1: a human cell line of endothelial origin. In Vitro Cell. Dev. Biol. 28A: 136-148, 1992. PubMed: 1371504

Gucev ZS, et al. Evidence for insulin-like growth factor (IGF)-independent transcriptional regulation of IGF binding protein-3 by growth hormone in SKHEP-1 human hepatocarcinoma cells. Endocrinology 138: 1464-1470, 1997. PubMed: 9075703

Turner BM, Turner VS. Secretion of alpha1-antitrypsin by an established human hepatoma cell line and by human/mouse hybrids. Somatic Cell Genet. 6: 1-14, 1980. PubMed: 6245472

Nucleotide (GenBank) : S40179 pancreatic cholesterol esterase homolog {exon 10} [human, SK-Hep-I cells, ATCC HTB 52, Genomic, 84 nt].