JCRB9004:HeLa

JCRB9004 [HeLa]

Profile:The first aneuploid, epitherial-like cell line(Feb. 1951) from human tissue and maintained continuously by serial cell culture.

Date accepted: 05/21/1985

Previous cell Number: atcc

Animal: human

Sex and Age: F :31-year-old

Race: Black

Scientific Name: Homo sapiens

Case history: carcinoma of cervix

Classification: tumor

Histological type of the cell: epitheloid carcinoma

Tissue prepared: cervix

Origin of tumor: cervix

History of the cell: ATCC-->JCRB

Medium: Eagle's minimal essential medium with non essential amino acids, Earle's BSS and 10 % fetal bovine serum.

Passage method: Cells harvested after treated with 0.02% EDTA and 0.25% trypsin.

CO₂ Concentration: 5 %

Cell no. at passage: split ratio=1/2 - 1/10

Life span: infinite

Morphology: epithelial-like

Characteristics: first establish cell line in vitro from human

Cell ID data: available

DNA Profile: D5S818:11,12
D13S317:12,13.3
D7S820:8,12
D16S539:9,10
vWA:16,18
TH01:7
Amelogenin:X
TPOX:8,12
CSF1PO:9,10

Established by Gey,Coffma,Kubicek

Deposited by JCRB

Special notice: Not regulated

Cell bank: NIHS(JCRB)

References: (1,2,3,4,5)

Comments: EV=1.000 against other HeLa cells, like HeLaP3, HeLa229, and etc. Prepared from CCL 2.

(* The format of the table was revised on Apr.25.2003.)

[ Picture(s) ]

Cells(1), Cells(2), Cells(3), Mycoplasma test, Mycoplasma (PCR), Isozyme analysis, STR-PCR(1), STR-PCR(2), STR-PCR(3), Growth Curve, Freezing process, Others(1), Others(2), Others(5), Others(6), Others(7), Others(8)

[ Extra document(s) ]

No extradocuments available.

Date downloaded:

[ Record(s) of stored ampules ]

[ Lot No.: 062185: token/seed (JCRB9004) ]

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Viability under microscope (%):

Tests for mycoplasma, bacteria, and fungi were negative.

Anchorage dependency:

Cell Identification:

RFLP data:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium with non essential amino acids and 10% fetal calf serum.

Freezing medium:

Culture medium with 5% DMSO.

Passage method:

0.02% EDTA and 0.25% Trypsin.

Growth temperature:

CO₂ concentration:

Cell No. at passage:

split ratio = 1/2 - 1/10

Additional comments:

Prepared at IFO from CCL 2. Viability 91% was checked at 01/30/89.

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 092286: distribution (JCRB9004) ]

Passage Number,

Date cell culture started:

Concentration of cells in an ampule:

Antibiotics used :

Tests for mycoplasma, bacteria, and fungi were negative.

Tested Isoenzymes

AST,G6PD,LD,MD,MPI,NP,PEPB.

[ Culture condition of this lot.]

Culture medium:

MEM w/ non essential amino acids + FCS 10%

Passage method:

0.02% EDTA and 0.25% Trypsin.

Growth temperature:

Cell No. at passage:

Additional comments:

Viability = 94%, P.E = 36% prior to freezing.

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 020689: distribution (JCRB9004) ]

Passage Number,

Date cell culture started:

Concentration of cells in an ampule:

Antibiotics used :

[ Culture condition of this lot.]

Culture medium:

MEM w/ non essential amino acids + FCS 10%

Passage method:

0.02% EDTA and 0.25% Trypsin.

Growth temperature:

Cell No. at passage:

Additional comments:

Viability was 98.8% prior to freezing.

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 100490: distribution (JCRB9004) ]

Passage Number,

Date cell culture started:

Concentration of cells in an ampule:

Antibiotics used :

[ Culture condition of this lot.]

Culture medium:

MEM w/ non essential amino acids + FCS 10%

Passage method:

0.02% EDTA and 0.25% Trypsin.

Growth temperature:

Cell No. at passage:

Additional comments:

Viability was 98.5% before freezing.

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 032592: distribution (JCRB9004) ]

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Saturated cell density:

Growth rate:

Plating efficiency:

40.2 %
Viability under microscope (%):

97.0
Antibiotics used :free

Tests for mycoplasma, bacteria, and fungi were negative.

[ Culture condition of this lot.]

Culture medium:

EMEM + non essential A.A + FBS 10%(lot;J.R.Scientific,DO 01400

Passage method:

0.25% trypsin + 0.02% EDTA

Growth temperature:

Cell No. at passage:

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 032593: distribution (JCRB9004) ]

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Growth rate:

Plating efficiency:

49.3%
Viability under microscope (%):

99.2
Antibiotics used :free

Tests for mycoplasma, bacteria, and fungi were negative.

Tested Isoenzymes

Anchorage dependency:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium (Nissui) with non-essential amino acids(GIBCO 100x) and 10% FBS

Passage method:

Wash cells once with PBS(-) and treat with 0.25%- trypsin and 0.02%-EDTA for 5 min. at 37 C.

Growth temperature:

Cell No. at passage:

1.23x10^5 cells/ml

[ Additional Data ]

(Date downloaded: 2004-08-17)

[ Lot No.: 071994: distribution (JCRB9004) ]

Cells are propagated from the stock,

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Growth rate:

Viability under microscope (%):

97.6
Antibiotics used :free

Tests for mycoplasma, bacteria, and fungi were negative.

Tested Isoenzymes

Anchorage dependency:

Cell Identification:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium with non-essential amino acids and 10% heat-inactivated FBS/0070893

Passage method:

Washed cells with PBS(-) and treated with 0.25% trypsin and 0.02% EDTA for 5-8 min. at 37 C.

Growth temperature:

Cell No. at passage:

2-4x10^4 cells/sq.cm

Memo:

[ Additional Data ]

STR-PCR(1),

STR-PCR(2),

(Date downloaded: 2004-08-17)

[ Lot No.: 051299: seed (JCRB9004) ]

Cells are propagated from the stock,

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Viability under microscope (%):

96.6
Antibiotics used :free

Tests for mycoplasma, bacteria, and fungi were negative.

Tested Isoenzymes

G6PD, LD, NP were examined and confirmed as HeLa..

Anchorage dependency:

Cell Identification:

RFLP data:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium with non-essential amino acids and 10% FBS (Intergen RB52405).

Freezing medium:

Culture medium with 5% DMSO.

Passage method:

Cells harvested with 0.25% trypsin and 0.02% EDTA.

Growth temperature:

CO₂ concentration:

Cell No. at passage:

2.5 x 10^4 cells/sq.cm.

Memo:

[ Additional Data ]

Mycoplasma test,

Mycoplasma (PCR),

STR-PCR(1),

STR-PCR(2),

STR-PCR(3),

Freezing process,

(Date downloaded: 2004-08-17)

[ Lot No.: 05232001: distribution (JCRB9004) ]

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Viability under microscope (%):

92
Antibiotics used :free

Tests for mycoplasma, bacteria, and fungi were negative.

Tested Isoenzymes

NP, LDH and G6PD(typeA) examined. Human, HeLa..

Anchorage dependency:

Cell Identification:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium with NEAA and 10% fetal bovine serum (Intergen RB52405).

Freezing medium:

Culture medium with 5% DMSO.

Passage method:

Cells harvested after treatment with 0.25% trypsin and 0.02% EDTA at room temperature for 5 minutes. Subculture once a week.

Growth temperature:

CO₂ concentration:

Cell No. at passage:

1.5 x 10^4 cells/sq.cm.

Additional comments:

Mycoplasma tested by PCR method.

[ Additional Data ]

Mycoplasma test,

Mycoplasma (PCR),

Isozyme analysis,

STR-PCR(1),

STR-PCR(2),

STR-PCR(3),

Freezing process,

(Date downloaded: 2004-08-17)

[ Lot No.: STD2004: research (JCRB9004) ]

Cells are propagated from the stock,

Passage Number,

Cell culture was started without cloning from the previous stock.

Date cell culture started:

Concentration of cells in an ampule:

Viability under microscope (%):

98.0
Antibiotics used :free

Sterility test has not completed yet.

Anchorage dependency:

Cell Identification:

[ Culture condition of this lot.]

Culture medium:

Eagle's minimal essential medium with non-essential amino acids and 10% fetal bovine serum (Intergen RB52305).

Freezing medium:

Culture medium containing 5% DMSO.

Passage method:

Cells were harvested after treatment with 0.25% trypsin and 0.02% EDTA at room temperature for three minutes.

Growth temperature:

CO₂ concentration:

Cell No. at passage:

6.0 x 10^4 cells/ml

[ Additional Data ]

STR-PCR(1),

STR-PCR(2),

STR-PCR(3),

Freezing process,

(Date downloaded: 2004-08-17)

Exported from cellroot.dbf and cellspec.dbf.

Reference List

1. Gey,G.O., Coffman,W.D., and Kubicek,M.T. Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium, Cancer Res., 12: 264-265, 1952.

2. Scherer,W.F., Syverton,J.T., and Gey,G.O. Studies on the propagation in vitro of poliomyelitis viruses, J. Exp. Med., 97: 695-710, 1953.

3. Salzman,N.P. Animal cell cultures: Tissue culture is a powerful tool in the study of nutrition, physiology, virology, and genetics., Science, 133: 1559-1565, 1961.

4. ATCC. CCL-2. In: R.Hay (ed.), <[13] ,Page Numbers>Rockville, MD USA: ATCC. 1994
. Notes: ATCC CCL-2, HeLa (Epitheloid carcinoma, cervix, human), 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. he 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). *[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.
Reference ID: 4091
5. Macville,M., Schrock,E., Padilla-Nash,H., Keck,C., Ghadimi,B.M., Zimonjic,D., Popescu,N., and Ried,T. Comprehensive and definitive molecular cytogenetic characterization of HeLa cells by spectral karyotyping, Cancer Res, 59: 141-150, 1999.
Notes: Genome Technology Branch, National Human Genome Research Institute, NIH, Bethesda, Maryland 20892, USA mmacville@patholaznnl
Abstract: We revisited the cytogenetic alterations of the cervical adenocarcinoma cell line HeLa through the use of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH). SKY analysis unequivocally characterized all abnormal chromosomes. Chromosomal breakpoints were primarily assigned by simultaneous assessment of SKY painted chromosomes and inverted 4,6-diamidino2-phenylindole banding from the same cell. Twenty clonally abnormal chromosomes were found. Comparison with previously reported HeLa G-banding karyotypes revealed a remarkably stable cytogenetic constitution because 18 of 20 markers that were found were present before. The classification of 12 markers was refined in this study. Our assignment of the remaining six markers was consistent with those described in the literature. The CGH map of chromosomal copy number gains and losses strikingly matched the SKY results and was, in a few instances, decisive for assigning breakpoints. The combined use of molecular cytogenetic methods SKY, CGH, and FISH with site-specific probes, in addition to inverted 4,6-diamidino-2-phenylindole or conventional G-banding analysis, provides the means to fully assess the genomic abnormalities in cancer cells. Human papillomaviruses (HPVs) are frequently integrated into the cellular DNA in cervical cancers. We mapped by FISH five HPV18 integration sites: three on normal chromosomes 8 at 8q24 and two on derivative chromosomes, der(5)t(5;22;8)(qll;q11q13;q24) and der(22)t(8; 22)(q24;q13), which have chromosome 8q24 material. An 8q24 copy number increase was detected by CGH. Dual-color FISH with a c-MYC probe mapping to 8q24 revealed colocalization with HPV18 at all integration sites, indicating that dispersion and amplification of the c-MYC gene sequences occurred after and was most likely triggered by the viral insertion at a single integration site. Numerical and structural chromosomal aberrations identified by SKY, genomic imbalances detected by CGH, as well as FISH localization of HPV18 integration at the c-MYC locus in HeLa cells are common and representative for advanced stage cervical cell carcinomas. The HeLa genome has been remarkably stable after years of continuous cultivation; therefore, the genetic alterations detected may have been present in the primary tumor and reflect events that are relevant to the development of cervical cancer
Reference ID: 5247