JCRB Cell Bank

[EHS (IFO50268): protocol for transplantation]

EHS is non cultivable cell line (does not grow in vitro). EHS can grow only by the transplantation to syngenic mice.

JCRB’s transplantation protocol detail.

1.	To remove DMSO, the suspension of tissue fragments was transfer to 10 mL of HBSS (or saline) in 50 mL tube(*1). 
2.	Centrifuge at 300 g for 5 min.
3.	Remove the supernatant.
4.	Re-suspend the tissue fragments in 10 mL of HBSS (or saline)
5.	Centrifuge at 300 g for 5 min.
6.	After the second centrifugation, remove the supernatant thoroughly with pipettman tip.

(*1) At the last step, the tissue fragments were aspirate by 2 mL syringe. So, use of 50 mL tube is adequate).
(*2) The reason for 2 times washing and use of pipettman tip is to remove the DMSO thoroughly.

7.	Re-suspend the tissue fragments to approximately 700 uL of saline. [The volume (700 uL) is larger than 200 uL x 2 mice (=400 uL), since the suspension is aspirated by syringes later.]

8.	(JCRB used pipetteman tip to resuspend the tissue fragment to loosen the tissue pellet.)
9.	Using 2 mL syringe x 2 with 18 G needle, aspirate the suspension of tissue fragments to 2 syringes (approx. 300 uL/syringe) (*3).

(*3) The reason why to prepare 2 syringes is to transplant the tissue suspension to 2 mice. If only one syringe is used for transplantation to multiple mice, there is a risk for clogging of tissues in the needle. In addition, it may cause an uneven amount of transplantation to each mouse.

10.	Transfer these syringes on ice(*4) to breeding room for experimental mice. 

(*4) JCRB’s procedure is to stand the syringe in 50 mL tubes cooled with ice.

11.	Change the needle to 20G
12.	Inject approximately 200 uL of tissue fragment suspension to one mouse subcutaneously(*5).

(*5) In some cases, the tissue fragments may be clogged in 20G needle. So, it is recommended to prepare 18G needle for the safe.

See also, 
https://cellbank.nibn.go.jp/~cellbank/images/pdf/IFO50268_EHS_001.pdf

[Close]

• 07282003
• 374
• 06302021
• 07192021
• 10202022
• 10272022

Cell No.	IFO50268	Cell Name	EHS
LOT No.	06302021	Lot Specification	distribution
Medium	Not cultivable. Subcutaneous transplantation (host; 10 week C57BL/6 strain mice).	Temperature
Cell Density at Seeding		Methods for Passages	Subcutaneous transplantation (neck). Tumors were removed from animals under sterile condition. (Tumor growth will take about 4 weeks.)
Doubling Time		Cell Number in Vial (cells/1ml)	uncountable
Viability at cell freezing (%)		Antibiotics Used
Passage Number		PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS in 1:1 mixture of EMEM and Hanks' BSS.	CO2 Conc.
Viability immediately after thawing (%)	uncountable	Additional information

Images

Cell No.	IFO50268	Cell Name	EHS
LOT No.	07192021	Lot Specification	distribution
Medium	Not cultivable. Subcutaneous transplantation (host; 10 week C57BL/6 strain mice).	Temperature
Cell Density at Seeding		Methods for Passages	Subcutaneous transplantation (neck). Tumors were removed from animals under sterile condition. (Tumor growth will take about 4 weeks.)
Doubling Time		Cell Number in Vial (cells/1ml)	uncountable
Viability at cell freezing (%)		Antibiotics Used
Passage Number		PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing	MEM with 20% FBS and10% DMSO	CO2 Conc.
Viability immediately after thawing (%)	uncountable	Additional information	Type of ampoules is plastic.

Images

• Contact us
• Privacy policy