-Detailed Information [JCRB1091]-

JCRB1091 NCE SVIIA3

Cell information

Important Notice(s)

Supplements for the Serum free culture medium

Supplemented growth hormones for the MCDB153 medium. 

Yasumoto,S. Kanagawa Cancer Center Research Institute. 

[Supplemented growth hormones for the MCDB153 medium.]

5 ug/ml Inslin(1% CH3COOH-DDW) 
0.5ug/ml Hidrocortisone(EtOH) 
10ug/ml transferrin(PBS) 
0.1mM phosphorylethanolamine(DDW) 
0.1mM ethanolamine(DDW) 
10ng/ml EGF(0.1% BSA-PBS) 
40-70ug/ml BPE(PBS) 

[Reference]
 
Jikken Igaku, Supplement
 BioManual Up series (2nd Edition)
 "New Cell Culture methods for Molecular Biology"
 T.Kuroki, N.Hoh and K.Chida
 Yohdo-sha,Co.
 Section 4. Primary cell culture, 2.Keratinocyte P71-80

[Medium]
MCDB153medium Powder (SIGMA cat.No.M-7403) for 1L and adjust to pH7.5 by adding Sodium Bicarbonate 1.176g(1N-NaOH).

MCDB153medium liquid (Functional Peptide Laboratory Inc., IFP) 1L and adust pH to 7.5 by adding stock solution of the sodium bicarbonate (x100-x1000).

[Working solution]
 
Store the working solution in the refregerator up to one month when supplements were added to the medium and only two weeks can be stored in the refregerator when BPE was added.

-- Medium -- 
MCDB153medium liquid (Functional Peptide Laboratory Inc., IFP) 1L and adjust pH to 7.5 by adding stock solution of the sodium bicarbonate (x100-x1000). 

-- Supplement--
Insulin(SIGMA cat.No.I-1882): 5mg/L (1000 x stock solution is prepared in 1% CH3COOH)
Hydrocortisone (SIGMA cat.No.H-4001): 0.5mg/L (10000 x stock solution is prepared in EtOH) 
Transferin (SIGMA cat.No.T8158): 10mg/L (1000 x stock solution is prepared in PBS)  
Ethanolamine (SIGMA cat.No.E-0135): 6.1mg/L (6 ul/L) 
Phosohorylethanolamine (SIGMA P-0503): 14.1mg/L 
EGF (SIGMA cat.No.E-4127): 10ug/L  (1000 x stock solution is prepared in 0.1% BSA-PBS )
BPE(Kyokuto Pherm. Co. cat.No.202000): 40-70mg/L (100 x stock solution is prepared in PBS)  

[Subculture] 

Trypsin solution: Trypsin typeIII(SIGMA cat.No.T-8253) 0.1%  0.02% EDTA in PBS- 
(SIGMA T-8253 was discontinued. Instead, Fujifilm Wako Cat. # 201-19181 will be adequate. Conventional 0.25% trypsin solution will also work.)
Trypsin Inhibitor solution: Trypsin Inhibitor(SIGMA cat.No.T-6522) 2mg/ml in PBS- 
 


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Cell type：general cells (View Pricing Information)

JCRB No.	JCRB1091	Cell Name	NCE SVIIA3
Profile	SV40 transformed keratinocyte cells derived from human cervix.	Other Name
Animal	human	Strain
Genus	Homo	Species	sapiens
Sex	F	Age	adult
Identity	available	Tissue for Primary Cancer	cervix
Case history		Metastasis
Tissue Metastasized		Genetics	SV40 ori(-), teromerase positive, P75NTR positive.
Life Span	infinite	Crisis PDL
Morphology	epithelial-like	Character	SV40 transformed human keratinocyte from cervix.
Classify	transformed	Established by	Yasumoto,S.
Registered by	Yasumoto,S.	Regulation for Distribution	Not apllied.
Comment		Year	2001
Medium	MCDB153 medium (0.03 mM Ca2+) with 5 ug/ml insulin, 0.5 ug/ml hydrocortisone, 10 ug/ml transferrin, 0.1 mM phosphorylethanolamine, 0.1 mM ethanolamine, 10 ng/ml EGF and 40-70 ug/ml bovine pituitary extract.	Methods for Passages	Cells are harvested with 0.1% trypsin and 0.1% EDTA. Medium change every 2 days and subculture every 10 days.
Cell Number on Passage	1.5 - 2.4 x 10^5 cells/sq.cm.	Race	Japanese
CO2 Conc.		Tissue Sampling	cervix
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_8374

Reference
Pubmed id:12747968	Gene expression of telomerase related proteins in human normal oral and ectocervical epithelial cells. Fujimoto R,Kamata N,Taki M,Yokoyama K,Tomonari M,Nagayama M,Yasumoto S Oral Oncol. 2003 Jul;39(5):445-52

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LOT Information

Viability/Growth rate/Cell number are represented as actual values measured at lot presentation in JCRB, but are not guaranteed values. Additionally, the doubling time is a rough value measured during passages.

11102005

Cell No.	JCRB1091	Cell Name
LOT No.	11102005	Lot Specification	distribution
Medium	MCDB153 medium with 5 ug/ml insulin, 0.5 ug/ml hydrocortisone, 10ug/ml transferrin, 0.1mM phosphoethanolamine, 0.1 mM ethanolamine, 10 ng/ml EGF, 50 ug/ml BPE (serum-free).	Temperature	37 C
Cell Density at Seeding	2.7 - 5.9 x 10^4 cells/ml	Methods for Passages	0.025% trypsin and 0.02% EDTA. Trypsin activity was stopped with soybean trypsin inhibitor.
Doubling Time	approx. 3 days	Cell Number in Vial (cells/1ml)	1.1 x 10^6
Viability at cell freezing (%)	54.3	Antibiotics Used	free
Passage Number	P22	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	Confirmed as human by NP, G6PD (type B), LD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)	D5S818:12 D13S317:11,12 D7S820:8,11 D16S539:11,12 VWA:16,18 TH01:7 AM:X TPOX:9,12 CSF1PO:11,12
Adhesion	Yes	Exoteric Gene
Medium for Freezing	10% DMSO, 0.1% methylcellulose - culture medium	CO2 Conc.	5%
Viability immediately after thawing (%)		Additional information

JCRB Cell Bank

JCRB1091 NCE SVIIA3

Cell information

LOT Information