HeLa (Epithelioid carcinoma, cervix, human)

ATCC CCL-2

Current medium for propagation: Eagle's MEM with non-essential amino acids and Earle's BSS, 90%; FBS, 10%. HeLa was the first aneuploid, epithelial-like cell line to be derived from human tissue and maintained continuously by serial cell culture. It was isolated by G.O. Gey, W.D. Coffman, and M.T. Kubicek in February, 1951, from a carcinoma of the cervix of a 31-year-old Black female (Cancer Res. 12: 264, 1952). In a recent re-examination of the original slides Jones et al. (Obstet. Gynecol. 38: 945-949, 1971) diagnosed the tumor as an adenocarcinoma. Since its origin, it has been one of the most widely studied cell lines. A plasma clot culture was sent to W.F. Scherer in May, 1952, who maintained it serially in monolayer cultures in a medium consisting of human serum, 40%; chick embryo extract, 2%; and Hanks' balanced salt solution, 58%. In 1954, the line was frozen (Proc. Soc. Exp. Biol. Med. 87: 480, 1954), and stored until 1959, at which time it was recultured and refrozen until 1960. After another recultivation and freezing, it was reactivated in November, 1961, for the American Type Culture Collection. The line was submitted at approximately the 76-88 passage level, and may be considered most similar in characteristics to the cells described in the classic studies of Scherer, Syverton, and Gey (J. Exp. Med. 97: 695, 1953). HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences (EMBO J. 3: 1151-1157, 1984; ibid., 5: 2285-2292, 1986; Am. J. Pathol. 119: 361-366, 1985; Nature 314: 111-114, 1985; Virology 145: 313-318, 1985; Proc. Natl. Acad. Sci. USA 88: 5523-5527, 1991). Handle as potentially biohazardous material under at least Biosafety Level 2 containment. DESCRIPTION OF REPOSITORY REFERENCE SEED STOCK Number of Serial Subcultures from Tissue of Origin: Unknown; 90-102 from culture received by W.F. Scherer, 1952. Freeze Medium: Basal medium (Eagle) with Hanks' BSS, 80%; human serum, 15%; glycerol, 5%; antibiotic-free. Viability: Approximately 90% (dye exclusion). Culture Medium: Minimum essential medium (Eagle) with non-essential amino acids and Earle's BSS, 90%; human serum, 10%; antibiotic-free. Growth Characteristics of Thawed Cells: An inoculum of 0.5-1.0 X 10(5) viable cells/ml in above culture medium at 37C, in an atmosphere of 5% carbon dioxide-95% air, multiplies approximately 15-fold in 7 days. Plating Efficiency: Approximately 45% in the above culture medium. Morphology: Epithelial-like. Karyology: Chromosome Frequency Distribution 50 Cells: 2n = 46 Cells: 1 5 2 5 9 13 8 3 1 1 1 1 Chromosomes: 70 78 79 80 81 82 83 84 85 86-147-164 There is a small telocentric chromosome in 98% of the cells. 100% aneuploidy in 1385 cells examined. Four typical HeLa marker chromosomes have been reported in the literature. M1 is a rearranged long arm and centromere of chromosome 1 and the long arm of chromosome 3. M2 is a combination of short arm of chromosome 3 and long arm of chromosome 5. M3 is an isochromosome of the short arm of chromosome 5. M4 consists of the long arm of chromosome 11 and an arm of chromosome 19. HeLa Marker Chromosomes: One copy of Ml, one copy of M2, four-five copies of M3, and two copies of M4 as revealed by G-banding patterns. Sterility: Tests for mycoplasma, bacteria, and fungi were negative. Species: Confirmed as human by immunofluorescence test. Virus Susceptibility: Susceptible to poliovirus type 1 and adenovirus type 3. Reverse Transcriptase: Not detected. Isoenzymes: G6PD type A. Submitted by: W.F. Scherer, Department of Microbiology, Cornell University School of Medicine, New York, NY. Prepared and characterized by: Child Research Center of Michigan, Detroit, MI, and ATCC, Rockville, MD. Revised: March 1994