-Detailed Information [JCRB4117]-

JCRB4117 NCC-CCS1B-X2-C1

Cell information

Important Notice(s)

Culture medium and substratum for NCC-series of sarcoma cell lines / NCCシリーズ肉腫細胞株の培地と培養基質

[Culture medium and substratum for NCC-series of sarcoma cell lines / NCCシリーズ肉腫細胞株の培地と培養基質]

NCC-series of sarcoma cell lines should be cultivated with DMEM/F12 with supplements as below.

[Complete medium]

DMEM/F12(with GLUTAMAX): Thermofisher (GIBCO) # 10565018
5% FBS
5 ug/mL recombinant human insulin: Fujifilm Wako # 093-06471
0.4 ug/mL hydrocortisone: Sigma # H4001
5 ng/mL recombinant human EGF: Sigma # E9644
10 ng/mL recombinant human basic FGF: Thermofisher (GIBCO) # PHG0369
5 uM Y-27632: Selleck # S1049

The addition of antibiotics and antimycotic is optional.

[Stock solution of supplements]

Insulin (1000X): 5 mg/mL in 5 mM HCl: (add 500 uL to 500 mL DMEM/F12)
Hydrocortisone (2500X): 1 mg/mL in EtOH: (add 200 uL to 500 mL DMEM/F12)
EGF (2000X): 10 ug/mL in 0.1% BSA/PBS: (add 250 uL to 500 mL DMEM/F12)
bFGF (1000X): 10 ug/mL in 0.1% BSA/PBS: (add 500 uL to 500 mL DMEM/F12)
Y-27632 (2000X): 10 mM in water: (add 250 uL to 500 mL DMEM/F12)

These stock solutions should be dispensed in small aliquots and preserved at -80C. You can mix these stock solutions and preserve as cocktail.

[Culture substrate]

Although non-coated tissue culture grade dish/flask was originally used for NCC-series of sarcoma cell lines, JCRB often used type I collagen-coated dish/flask for the cultivation of these cell lines since the adhesion of cells was sometimes very poor. 

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NCCシリーズ肉腫由来細胞株の分譲について：営利機関への分譲には細胞樹立機関である国立がん研究センターとの有体物提供契約書の締結が必要です

[NCCシリーズ肉腫由来細胞株の分譲について：営利機関への分譲には細胞樹立機関である国立がん研究センターとの有体物提供契約書の締結が必要です]

NCCシリーズ肉腫由来細胞株の営利機関への分譲においては、当該営利機関と国立がん研究センターとの間で別途有償の有体物提供契約書の締結が条件とされています。

当該細胞の分譲をご希望される営利機関は分譲の依頼に先立ち、下記、細胞樹立機関の近藤格先生および国立がん研究センター産学連携・知財戦略課へ連絡のうえ、有体物提供契約書の締結後、そのコピー（契約成立を証明する箇所で可）を添付してJCRB細胞バンクに分譲依頼を行ってください。

なお、連絡を取る際に、ＪＣＲＢ番号、細胞名、研究（使用）目的を明確にお伝えください。

連絡先：
〒104-0045東京都中央区築地5－1－1
国立研究開発法人国立がん研究センター 研究所 希少がん研究分野
近藤 格
メールアドレス： takondo[at]ncc.go.jp    *[at]を@に置き換えてください。

〒104-0045東京都中央区築地5－1－1
国立研究開発法人国立がん研究センター産学連携・知財戦略課
メールアドレス： patent[at]ml.res.ncc.go.jp    *[at]を@に置き換えてください。

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Conclusion of Material Transfer Agreement with charge (MTA) with National Cancer Center (NCC) is prerequisite for the transfer of NCC-series of sarcoma-derived cell lines to For-profit Organizations.

[Conclusion of Material Transfer Agreement with charge (MTA) with National Cancer Center (NCC) is prerequisite for the transfer of NCC-series of sarcoma-derived cell lines to For-profit Organizations]

The transfer of NCC-series of sarcoma-derived cell lines to For-profit Organizations requires the conclusion of MTA with NCC.

Prior to the order to JCRB Cell Bank, For-profit Organizations need contacts with Dr.Tadashi KONDO of NCC (the cell lines establisher) and Research Alliance and Intellectual Property Division of NCC for the conclusion of MTA.  At your contact, please clarify the JCRB number, name of cell line and intended use of cells.

Order to JCRB Cell Bank must be accompanied with the copy of the part of MTA that certifies the conclusion of MTA, together with Request and Agreement Form (Form A).

Contact Information:

Address: 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045 Japan
National Cancer Center Research Institute
Division of Rare Cancer Research
Dr. Tadashi KONDO
E-mail: takondo[at]ncc.go.jp          *[at] to be replaced to @.

Address: 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045 Japan
National Cancer Center Research Alliance and Intellectual Property Division
E-mail: patent[at]ml.res.ncc.go.jp       *[at] to be replaced to @.

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Cell type：general cells (View Pricing Information)

JCRB No.	JCRB4117	Cell Name	NCC-CCS1B-X2-C1
Profile	Clear cell sarcoma cell line (Established from a second generation xenograft established by subcutaneous injection in female NOD/Shi-scid IL-2R gamma null mice)	Other Name
Animal	human	Strain
Genus	Homo	Species	sapiens
Sex	F	Age	43-year-old
Identity	available	Tissue for Primary Cancer	Left thigh
Case history	clear cell sarcoma	Metastasis
Tissue Metastasized		Genetics	EWSR1-ATF1(fusion)
Life Span	Not confirmed	Crisis PDL
Morphology	Spindle-shape	Character	doubling time: 20 hr (Original description. D.T. = 1.5-2 days in JCRB's culture.)
Classify	tumor	Established by	Marimu SAKAMOTO
Registered by	Tadashi KONDO	Regulation for Distribution	Additional MTA (for profit organization)
Comment		Year	2024
Medium	Original medium: DMEM with 10% FBS. (Current medium for propagation: DMEM/F-12 medium supplemented with GlutaMAX, 5% heat-inactivated fetal bovine serum (FBS), 10 uM Y-27632, 10 ng/mL basic fibroblast growth factor (bFGF), 5 ng/mL epidermal growth factor (EGF), 5 ug/mL insulin, 0.4 ug/mL hydrocortisone.)	Methods for Passages	0.1% trypsin-EDTA
Cell Number on Passage		Race	Japanese
CO2 Conc.	5%	Tissue Sampling	Left thigh
Tissue Type

Detection of virus genome fragment by Real-time PCR
Detected DNA Virus	not tested				Detected RNA Virus	not tested

Reference
Pubmed id:29197033	Establishment and proteomic characterization of patient-derived clear cell sarcoma xenografts and cell lines. Sakumoto M,Oyama R,Takahashi M,Takai Y,Kito F,Shiozawa K,Qiao Z,Endo M,Yoshida A,Kawai A,Kondo T In Vitro Cell Dev Biol Anim. 2018 Feb;54(2):163-176

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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.

01232026

Cell No.	JCRB4117	Cell Name	NCC-CCS1B-X2-C1
LOT No.	01232026	Lot Specification	distribution
Medium	DMEM/F12 supplemented with GlutaMax(GIBCO #10565018) with 5% heat-inactivated fetal bovine serum (Nichirei Cat. # 174112, Lot 20M00K), 5 ug/mL insulin, 0.4 ug/mL hydrocortisone, 5 ng/mL EGF, 10 ng/mL bFGF and 5 uM Y-27632.　Collagen coat plate used.	Temperature	37 C
Cell Density at Seeding	0.211-1.11x10^5 cells/sq.cm	Methods for Passages	Cells were harvested after treatment with 0.25%trypsin(GIBCO) and 0.02% EDTA.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	3.18x10^6
Viability at cell freezing (%)	92.2	Antibiotics Used	free
Passage Number	p34	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 D13S317:8 D7S820:11,12 D16S539:11 VWA:15,16 TH01:6 AM:X TPOX:11 CSF1PO:10
Adhesion	Yes	Exoteric Gene	NT
Medium for Freezing	BAMBANKER (LYMPHOTEC Inc., CS-02-001, NIPPON Genetics Co., LTD)	CO2 Conc.	5%
Viability immediately after thawing (%)	86.7	Additional information

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