-Detailed Information [JCRB0149]-

JCRB0149 Bhas 42

Cell information

Important Notice(s)

[問題が該当する細胞株]
JCRB0601 BALB/3T3 A31-1-1
IFO50070 Balb/3T3-A31-1-1
IFO50299 Balb/c 3T3 A31-1-13
IFO50298 Balb/c 3T3 A31-I-1
JCRB0149 Bhas 42
JCRB1355 1-1ras1000
JCRB1356 A31-1-1
JCRB1357 1-1 src
	
これらの細胞は、マウス系統解析から、Swissマウスに由来する細胞であり、Balb/c 3T3 clone A31に由来する細胞ではないことが判明しました。

Balb/c 3T3 A31-1-1, Balb/c 3T3 A31-1-13細胞は、親株をBalb/c 3T3 clone A31細胞とするサブクローンとして、角永先生の研究室（当時National Cancer Intitute）で樹立されました。これらのサブクローンは、親株のA31よりも化学発癌物質によるトランスフォーメーションに適するものとされ、世界的に広く利用されています。JCRB細胞バンクにも複数のルートから寄託されました。なお、IFO50070番で登録された株は、角永先生から直接バンクに寄託されたものです。
その後、さらに他の研究室によりBalb/c 3T3 A31-1-1を親株として遺伝子導入された細胞株、Bhas 42, 1-1ras1000, 1-1 srcが樹立され、JCRB細胞バンクに寄託されました。

しかし、JCRB細胞バンクが細胞の由来を確かめる目的で、これらの細胞株のマウス系統解析を行ったところ、角永先生の研究室で樹立されたこれらのサブクローン、および、A31-1-1に派生する細胞株はすべてSwissマウス由来であることがわかりました。

一方、元々の親株とされるBalb/c 3T3 clone A31細胞は、正しくBalb/cマウス由来であることが確認されました。

角永先生はすでにかなり前に逝去されており、どのような状況でこのような事態が生じたのかは確認ができませんが、Balb/c 3T3 A31-1-1 A31-1-13は、何らかの混同、あるいは誤認によりSwissマウスの線維芽細胞をクローンとして拾って樹立した株ということになります。

これらの細胞株は、歴史的にも広く用いられてきた経緯、あるいは試験法に記載されてきた経緯等があり、その有用性は失われておりません。このため、JCRB細胞バンクでは、これらの細胞の登録を抹消せず、分譲を継続しています。しかし、上記細胞の使用に関しては、Balb/c 3T3 clone A31細胞とは系統が異なる細胞であることをご了解ください。

例えば、もし、枝番がついていないA31細胞としての性質を求めるのであれば（たとえばOECDの光毒性試験など)、JCRB9005で登録されているBALB/3T3 clone A31のご利用をご検討ください。

一方、角永らにより樹立された、トランスフォームしやすいとされる細胞のご利用をお考えの場合、または、論文の追試や、試験法などで細胞名が明記されている場合には、マウス系統の問題はあるものの、その使用は妥当となります。

つきましては、細胞のご研究への適合性についてご留意くださいますようお願い申し上げます。

参考文献：In Vitro Cell Dev Biol Anim. http://link.springer.com/article/10.1007%2Fs11626-016-0104-3


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The problem of origin of Balb/c 3T3 A31-1 derived cell lines.

[The cell lines in problem]

JCRB0601 BALB/3T3 A31-1-1
IFO50070 Balb/3T3-A31-1-1
IFO50299 Balb/c 3T3 A31-1-13
IFO50298 Balb/c 3T3 A31-I-1
JCRB0149 Bhas 42
JCRB1355 1-1ras1000
JCRB1356 A31-1-1
JCRB1357 1-1 src

These cell lines have been proved to be derived from Swiss mouse cells, but not from the derivative of Balb/c 3T3 clone A31.

Balb/3T3 A31-1-1 and A31-1-13 were reported to be isolated from Balb/3T3 clone A31 by Dr. Kakunaga at National Cancer Institute in 1980 (Science 209: 505-507, 1980). These cell lines were reported to be useful for the study of chemically induced transformation of cells, and were widely used in the world. These cell lines were deposited at JCRB Cell Bank from different deposition routes. For example, IFO50070 strain was the one that was directly deposited by Dr. Kakunaga.

Thereafter, several laboratories developed genetically modified cell lines from Balb/c 3T3 A31-1-1. These are Bhas 42, 1-1ras1000, 1-1 src, and were also deposited at JCRB Cell Bank.

JCRB Cell Bank took place an analysis of mouse strain to check the origin of these cell lines. During this analysis, we found that A31-1-1, A31-1-13 and all these derivatives are NOT derived from Balb/c mouse, but from Swiss mouse.

Wheras, the parental cell line, Balb/c 3T3 clone A31 was confirmed as Balb/c mouse origin.

Dr. Kakunaga passed away in 1988, and unfortunately we can not trace the situation and cause of problem on this matter. However, it is suspected that the Balb/3T3 A31-1-1 and A31-1-13 subclones used in his laboratory was mistakenly establihed (cloned) from Swiss mouse-derived fibroblasts.

These cell lines have been historically used in worldwide, or have been described in test guidelines, and therefore the still valuable for research and testing. For this reason, JCRB does not exclude these cell lines from catalog and continues distribution of them. However, please keep in mind that these cell lines are not Balb/3T3 A31-derived cells.

For example, If your research needs the characteristics of Balb/c 3T3 clone A31 (such as OECE test guideline "In vitro 3T3 NRU phototoxicity test"), please consider to use JCRB9005 BALB/3T3 clone A31.

Wheras if you will use the cells established by Kakunaga for transformation studies, or if the name of cell lines are clearly described in the paper or test guidelines, these cell lines are still available for use regardless to the mouse strain problem.

Sincerely,

JCRB Cell Bank

References: In Vitro Cell Dev Biol Anim. Http://link.springer.com/article/10.1007%2 Fs 11626-016-0104-3
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Cell type：general cells (View Pricing Information)

JCRB No.	JCRB0149	Cell Name	Bhas 42
Profile	BALB 3T3 A31-1-1 cell line transfected with H-ras gene. The cell line keeps characteristics of the contact inhibition and the transfomation is induced by phorbol ester alone. No initialization required. (Strain problem was found. Refer Important Notice at cell detail page.)	Other Name
Animal	mouse	Strain	Swiss
Genus	Mus	Species	musclus
Sex		Age	embryo
Identity		Tissue for Primary Cancer	embryo
Case history	normal	Metastasis
Tissue Metastasized		Genetics	H-ras gene transfected BALB/c 3T3 A31-1-1 cell line. The origin of BALB/c 3T3 A31-1 was found not to be derived from BALB 3T3 A31.
Life Span	infinite	Crisis PDL
Morphology	fibroblast-like	Character	Growth on the dish is inhibited by cell to cell contact. Phorbol ester alone is enough to induce cellular transformation.
Classify	transfected	Established by	Sasaki,K.
Registered by	Sasaki,K.	Regulation for Distribution	free
Comment	Appearing of naturally occured transformation depends on serums and passage methods.	Year	2000
Medium	Eagle's minimal essential medium with 10% fetal bovine serum.	Methods for Passages	Cells are harvested after treatment with 0.25% trypsin and 0.02% EDTA. Split ratio is practically 1/100. Harvest cells before confluent at subculture.
Cell Number on Passage	2 x 10^4 cells/100mm dish.	Race
CO2 Conc.	5 %	Tissue Sampling	whole body of embryo
Tissue Type

Detection of virus genome fragment by Real-time PCR
Detected DNA Virus	tested				Detected RNA Virus	not tested
	CMV	-	parvoB19	-		-/negative. +/positive. nt/not tested. (positive (+) does not immediately mean the production of infectious viral particles.)
	EBV	-	HBV	-
	HHV6	-	HTLV-1	-
	HHV7	-	HTLV-2	-
	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_5494

Reference
Pubmed id:14729374	An assay method for the prediction of tumor promoting potential of chemicals by the use of Bhas 42 cells. Ohmori K,Sasaki K,Asada S,Tanaka N,Umeda M Mutat Res. 2004 Feb 14;557(2):191-202
Pubmed id:8481899	Tumor necrosis factor acts as a tumor promoter in BALB/3T3 cell transformation. Komori A,Yatsunami J,Suganuma M,Okabe S,Abe S,Sakai A,Sasaki K,Fujiki H Cancer Res. 1993 May 1;53(9):1982-5
Pubmed id:20702251	Transformation of ras transfected BALB 3T3 clone (Bhas 42) by promoters: Application for screening and specificity of promoters. Sasaki K,Mizusawa H,Ishidate M,Tanaka N Toxicol In Vitro. 1990;4(4-5):657-9
Pubmed id:3141328	Isolation and characterization of ras-transfected BALB/3T3 clone showing morphological transformation by 12-O-tetradecanoyl-phorbol-13-acetate. Sasaki K,Mizusawa H,Ishidate M Jpn J Cancer Res. 1988 Aug;79(8):921-30

LOT Information

11142013

Cell No.	JCRB0149	Cell Name	Bhas 42
LOT No.	11142013	Lot Specification	distribution
Medium	Eagle's minimal essential medium with 10% fetal bovine serum (FBS; GIBCO Cat # 10099).	Temperature	37C
Cell Density at Seeding	1.2 - 3.2 x 10^4 cells/mL	Methods for Passages	Cells were harvested after treatment with 0.25% trypsin and 0.02% EDTA. Split ratio is practically 1/100. Harvest cells before confluent at subculture.
Doubling Time		Cell Number in Vial (cells/1ml)	2.3 x 10^6 (cells/1 mL)
Viability at cell freezing (%)		Antibiotics Used	free
Passage Number	P15	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	G6PD,NP,LDH were examined. Mouse confirmed.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.	5%
Viability immediately after thawing (%)	96.7	Additional information