-Detailed Information [JCRB1318]-

[Information for genetic modification in HeLa9903 cell line]

The HeLa9903 is HeLa cell line that is stably expressing human estrogen receptor-alpha, reportor gene (estrogen response element linked to luciferase), and firefly luciferase gene (pGL3(Luc+)).
No viral vector was used for the introduction of external genes

[Information of transfection of external genes]

Unfortunately, the academic paper describing HeLa9903 has not been published. According to the patent document by the developer, Sumitomo Chemical Co. Ltd., the plasmid constructions and the method  will be as follows.

The introduced plasmids are

pGL3-TATA-EREx5-BSD: 
---> reportor gene (upstream sequences of Xenopus-derived vitronectin gene containing estrogen responce element was linked to firefly luciferase gene (Luc+))

pRC/RSV-hERalpha Kozak: 
---> human estrogen receptor alpha gene driven by RSV promotor.

At the transfection, these plasmids were digested with sal I, and linear DNAs were transfected to HeLa cells with lipofectamine.

The selection markers are, blasticidin S and neomycin (G418) resistant, respectively.
The developer used 16 ug/mL blasticidin S and 800 ug/mL G418 for the first selection.

The HeLa9903 cell line was deposieted at JCRB Cell Bank as stable transfectant, and JCRB cultivated the cells without the addition of these reagents. JCRB has not tried (tested) the above antibiotics selection.


[Close]

JCRB No.	JCRB1318	Cell Name	HeLa9903
Profile	hERalpha-HeLa-9903, transfected with estrogen receptor-alpha - reporter gene construct.	Other Name
Animal	human	Strain
Genus	Homo	Species	sapiens
Sex		Age
Identity	available	Tissue for Primary Cancer	uterine cervix
Case history		Metastasis
Tissue Metastasized		Genetics
Life Span		Crisis PDL
Morphology		Character
Classify		Established by
Registered by		Regulation for Distribution
Comment	HeLa clone.	Year	2008
Medium		Methods for Passages
Cell Number on Passage		Race	Black
CO2 Conc.		Tissue Sampling
Tissue Type

Detection of virus genome fragment by Real-time PCR
Detected DNA Virus	tested	Detected RNA Virus	tested
CMV	-	parvoB19	-	HCV	-	HTLV-1	-
EBV	-	HBV	-	HIV-1	-	HTLV-2	-
HHV6	-	HTLV-1	-	HIV-2	-	HAV	-
HHV7	-	HTLV-2	-	-/negative. +/positive. nt/not tested. (positive (+) does not immediately mean the production of infectious viral particles.)
BKV	-	HIV-1	-
JCV	-	HIV-2	-
ADV	-	HPV18	+
Notes

Link
Cellosaurus(ExPASy) Cellosaurus (ExPASy) is developed by Amos Bairoch of the CALIPHO group at the SIB - Swiss Institute of Bioinformatics as part of the neXtProt project. Cellosaurus is a knowledge database of cell lines with various information summarized.	CVCL_2485

Reference
Pubmed id:12001993	HeLa cells 50 years on: the good, the bad and the ugly. Masters JR Nat Rev Cancer. 2002 Apr;2(4):315-9
Pubmed id:16589695	A RAPID METHOD FOR VIABLE CELL TITRATION AND CLONE PRODUCTION WITH HELA CELLS IN TISSUE CULTURE: THE USE OF X-IRRADIATED CELLS TO SUPPLY CONDITIONING FACTORS. Puck TT,Marcus PI Proc Natl Acad Sci U S A. 1955 Jul 15;41(7):432-7

12062010
01292010
08242008

Cell No.	JCRB1318	Cell Name	HeLa9903
LOT No.	12062010	Lot Specification	distribution
Medium	DMEM(GIBCO) with 10% heat inactivated fetal bovine serum(SIGMA027K03911)	Temperature	37 C
Cell Density at Seeding	0.5-1.5x10^4 cells/sq.cm	Methods for Passages	Cells were harvested after treatment with 0.25%(GIBCO) trypsin and 0.02% EDTA.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.18x10^6
Viability at cell freezing (%)	95.8	Antibiotics Used	free
Passage Number	p4*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)	D5S818:11,12 D13S317:12,13.3 D7S820:8,12 D16S539:9,10 VWA:16,18 TH01:7 AM:X TPOX:8,12 CSF1PO:9,10
Adhesion	Yes	Exoteric Gene	NT
Medium for Freezing	Cell Banker BLC-1(Nihon Zenyaku Industries)	CO2 Conc.	5 %
Viability immediately after thawing (%)		Additional information

Images

Cell No.	JCRB1318	Cell Name
LOT No.	01292010	Lot Specification	distribution
Medium	Eagle's minimum essential medium (phenol red-free) with 10% charcol/dextran treated fetal bovine serum (Hyclone Cat. # SH30068.2)	Temperature	37 C
Cell Density at Seeding	5 - 10 x 10^4 cells/ml	Methods for Passages	Cells were harvested after treatment with 0.25% trypsin and 0.02% EDTA.
Doubling Time	approx. 33 hrs	Cell Number in Vial (cells/1ml)	1.3 x 10^6
Viability at cell freezing (%)	94.9	Antibiotics Used	free
Passage Number	Unknown (5 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	Confirmed as human by NP, G6PD (type A), MD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	Yes	Exoteric Gene
Medium for Freezing	10% DMSO, 20% charcol/dextran treated FBS - EMEM (phenol red-free)	CO2 Conc.	5%
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB1318	Cell Name	HeLa9903
LOT No.	08242008	Lot Specification	distribution
Medium	DMEM(GIBCO) with 10% heat inactivated fetal bovine serum(Hyclone GRD0051)	Temperature	37 C
Cell Density at Seeding	3.9x10^3 cells/sq.cm	Methods for Passages	Cells were harvested after treatment with 0.25%(GIBCO) trypsin and 0.02% EDTA.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	1.1x10^6
Viability at cell freezing (%)	94.9	Antibiotics Used	free
Passage Number	p3*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)	D5S818:11,12 D13S317:12,13.3 D7S820:8,12 D16S539:9,10 VWA:16,18 TH01:7 AM:X TPOX:8,12 CSF1PO:9,10
Adhesion	Yes	Exoteric Gene	NT
Medium for Freezing	Cell Banker BLC-1(Nihon Zenyaku Industries)	CO2 Conc.	5 %
Viability immediately after thawing (%)		Additional information

Images

JCRB Cell Bank

JCRB1318 HeLa9903

Cell information

LOT Information