-Detailed Information [JCRB1093]-

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- 
 


[Close]

JCRB No.	JCRB1093	Cell Name	PSVK1
Profile	Human normal keratinocyte prepared from human foreskin that was immortalized by the infection of the pSV40 ori-(minus) DNA.	Other Name
Animal	human	Strain
Genus	Homo	Species	sapiens
Sex	M	Age	0-year-old
Identity	available	Tissue for Primary Cancer	foreskin
Case history	normal	Metastasis
Tissue Metastasized		Genetics	Cells transfected by SV40 ori- (minus) DNA. Teromerase activity is maintained.
Life Span	infinite	Crisis PDL
Morphology	epithelial-like	Character	Human primary cell line immortalized by pSV40 gene.
Classify	transformed	Established by	Yasumoto,S.
Registered by	Yasumoto,S.	Regulation for Distribution	not applied
Comment		Year	2000
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. (serum-free)	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
CO2 Conc.		Tissue Sampling	foreskin
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_8403

Reference
Pubmed id:none	Telomere dynamics and telomerase activities during immortalization of human epithelial cells transfected with HPV16 and SV40 oncogenes. Ohji,H. Yokohama Igaku, 49: 35-43, 1998.

01052005
11262014
10272016
02282022

Cell No.	JCRB1093	Cell Name	PSVK1
LOT No.	01052005	Lot Specification	distribution
Medium	MCDB153 medium 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. (serum-free)	Temperature	37 C
Cell Density at Seeding	1.0 x 10^4/sq. cm	Methods for Passages	Cells harvested after treatment with 0.1% trypsin and 0.1% EDTA. Subculture once a week, medium change every 2-3 days.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.77x10^6
Viability at cell freezing (%)	97.2	Antibiotics Used	free
Passage Number	P32	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)	D5S818:12,13 D13S317:12 D7S820:10,11 D16S539:9,13 VWA:16 TH01:6,7 AM:X,Y TPOX:8,11 CSF1PO:11,12
Adhesion	Yes	Exoteric Gene	NT
Medium for Freezing	One to one mix with Cultue medium and FM-1(containing 10%DMSO) medium.	CO2 Conc.	5%
Viability immediately after thawing (%)		Additional information

Images

Cell No.	JCRB1093	Cell Name	PSVK1
LOT No.	11262014	Lot Specification	distribution
Medium	KGM-Gold (Lonza Cat.# 00192060). Originally the instructed medium was MCDB153 medium 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 BPE.	Temperature	37 C
Cell Density at Seeding	approx. 2 x 10^5 cells/mL	Methods for Passages	Cells are harvested with 0.1% trypsin and 0.02% EDTA. Trypsin activity was terminated by 0.1% (final conc.) soybean trypsin inhibitor (Sigma type IS) after trypsin treatment.
Doubling Time		Cell Number in Vial (cells/1ml)	1.9 x 10^6
Viability at cell freezing (%)	93.2	Antibiotics Used	free
Passage Number	P36	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	Confirmed as human by NP, G5PD (type B), MD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	Yes	Exoteric Gene
Medium for Freezing	0.1% methylcellulose, 10% DMSO - KGM-Gold.	CO2 Conc.	5%
Viability immediately after thawing (%)	93.2	Additional information

Cell No.	JCRB1093	Cell Name	PSVK1
LOT No.	10272016	Lot Specification	distribution
Medium	KGM-Gold (Lonza)	Temperature	37 C
Cell Density at Seeding	1.2 - 1.6 x 10^5 cells/mL	Methods for Passages	Cells are harvested with 0.1% trypsin and 0.1% EDTA. Trypsin action was terminated by 0.1% soybean trypsin inhibitor (Sigma, type IS).
Doubling Time	59 - 126 hrs.	Cell Number in Vial (cells/1ml)	3.4 x 10^6
Viability at cell freezing (%)	89.7	Antibiotics Used	free
Passage Number	P35	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	Yes	Exoteric Gene
Medium for Freezing	10% DMSO, 0.1% methylcellurose in culture medium	CO2 Conc.	5%
Viability immediately after thawing (%)	89.7	Additional information

Cell No.	JCRB1093	Cell Name	PSVK1
LOT No.	02282022	Lot Specification	distribution
Medium	KGM-Gold (Lonza)	Temperature	37 C
Cell Density at Seeding	0.6 - 3.5 x 10^5 cells/mL	Methods for Passages	Cells were harvested after treatment with 0.1% trypsin. Trypsin action was terminated by 0.1% soybean trypsin inhibitor (Sigma, type IS).
Doubling Time	35 - 82 hrs	Cell Number in Vial (cells/1ml)	2.5 x 10^6
Viability at cell freezing (%)	88.5	Antibiotics Used	free
Passage Number	P35	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	Yes	Exoteric Gene
Medium for Freezing	10% DMSO, 0.1% methylcellurose in KGM-Gold	CO2 Conc.	5%
Viability immediately after thawing (%)	86.8	Additional information

JCRB Cell Bank

JCRB1093 PSVK1

Cell information

LOT Information