SiHa (ATCC® HTB-35)

Organism: Homo sapiens, human  /  Tissue: cervix  /  Disease: grade II,squamous cell carcinoma

Organism Homo sapiens, human
Tissue
cervix
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain human papilloma virus
Disease grade II,squamous cell carcinoma
Age 55 years adult
Gender female
Ethnicity Asian
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 69; range = 51 to 72.
This is a hypertriploid human cell line with the modal chromosome number of 71, occurring in 24% of cells. Most cells had the chromosome numbers distributed between 69 and 72. Polyploid cells occurred at 7.6%. Fifteen or more marker chromosomes were common to most cells. Among them are dup(2) (q22q31) and del(2) (q31) which probably resulted from the balanced translocation between two N2s. Most cells had two copies of del(2). M2 is an A3-sized acrocentric. M13 is a minute submetacentric with 1-3 copies per cell. Origins of both M2 and M13 are not identified. There were two copies of normal X chromosomes. N2 was absent and probably was replaced by dup(2) and del(2).
Derivation
This line was established from fragments of a primary tissue sample obtained after surgery from a Japanese patient.
Clinical Data
55 years adult
Asian
female
This line was established from fragments of a primary tissue sample obtained after surgery from a Japanese patient.
Oncogene p53 +; pRB +
Genes Expressed
Oncogenes: p53 +; pRB +
Tumorigenic Yes
Effects
Yes, in nude mice; forms poorly differentiated epidermoid carcinoma (grade III)
Comments
Electron microscopic observations revealed presence of typical desmosomes at the cell junctions and an abundance of tonofilaments in the cytoplasm.
Mycoplasma contamination was detected and eliminated in 1975.
The line is reported to contain an integrated human papillomavirus type 16 genome (HPV-16, 1 to 2 copies per cell).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 12
D13S317: 11
D16S539: 12
D5S818: 9
D7S820: 10
THO1: 6,9
TPOX: 8
vWA: 14,17
Isoenzymes
AK-1, 1
ES-D, 2
G6PD, B
GLO-I, 2
Me-2, 1
PGM1, 1
PGM3, 1
Name of Depositor Y Ito
References

Baker CC, et al. Structural and transcriptional analysis of human papillomavirus type 16 sequences in cervical carcinoma cell lines. J. Virol. 61: 962-971, 1987. PubMed: 3029430

Pater MM, Pater A. Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix. Virology 145: 313-318, 1985. PubMed: 2992153

Yee C, et al. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am. J. Pathol. 119: 361-366, 1985. PubMed: 2990217

Friedl F, et al. Studies on a new human cell line (SiHa) derived from carcinoma of uterus. I. Its establishment and morphology. Proc. Soc. Exp. Biol. Med. 135: 543-545, 1970. PubMed: 5529598

Scheffner M, et al. The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines. Proc. Natl. Acad. Sci. USA 88: 5523-5527, 1991. PubMed: 1648218

Hendricks DT, et al. FHIT gene expression in human ovarian, endometrial, and cervical cancer cell lines. Cancer Res. 57: 2112-2115, 1997. PubMed: 9187105

Olive PL, Banath JP. Multicell spheroid response to drugs predicted with the comet assay. Cancer Res. 57: 5528-5533, 1997. PubMed: 9407963

Cross References

Nucleotide (GenBank) : AF001600 Human papillomavirus type 16 integrated SiHa HPV-16 variant DNA, capsid protein gene, complete cds, and flanking cellular DNA.

Nucleotide (GenBank) : AF001599 Human papillomavirus type 16 integrated SiHa HPV-16 variant, replication protein gene, complete cds, and flanking cellular sequence.

Basic Documentation
Other Documentation
References

Baker CC, et al. Structural and transcriptional analysis of human papillomavirus type 16 sequences in cervical carcinoma cell lines. J. Virol. 61: 962-971, 1987. PubMed: 3029430

Pater MM, Pater A. Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix. Virology 145: 313-318, 1985. PubMed: 2992153

Yee C, et al. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am. J. Pathol. 119: 361-366, 1985. PubMed: 2990217

Friedl F, et al. Studies on a new human cell line (SiHa) derived from carcinoma of uterus. I. Its establishment and morphology. Proc. Soc. Exp. Biol. Med. 135: 543-545, 1970. PubMed: 5529598

Scheffner M, et al. The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines. Proc. Natl. Acad. Sci. USA 88: 5523-5527, 1991. PubMed: 1648218

Hendricks DT, et al. FHIT gene expression in human ovarian, endometrial, and cervical cancer cell lines. Cancer Res. 57: 2112-2115, 1997. PubMed: 9187105

Olive PL, Banath JP. Multicell spheroid response to drugs predicted with the comet assay. Cancer Res. 57: 5528-5533, 1997. PubMed: 9407963