The clonal human embryonic progenitor cell line RASMO12 was derived from the registered parental hES cell line H9 (WA09) as described (West et al, 2008). It expresses mark­ers of intermediate mesoderm (Lusis et al, 2010) including: Wilms tumor 1 gene (WT1), nephronectin (NPNT), TSHZ2, SIX1, and distal HOX gene expression HOXA4, B8, C8. Upon differentiation with kit ES-K10, the line expresses the neu­rotransmitter (GABA) transporter SLC6A12 (Rasola et al, 1995) and SALL1.

Positive mRNA Markers: WT1, NPNT, TSHZ2, SLC6A12, SALL1, SIX1

Product Information:

Derivation: Progenitor line W10 was derived from the parental NIH registered human ES cell line H9 (WA09) as previously described (West, M.D. et al, 2008).

Cell Number: Vial contains >500,000 cells cryopreserved in 1 ml of FBS with 10% DMSO.

Recommended Growth Medium: ES-M181

Recommended Culture Conditions: Cells should be plated onto tissue culture grade polystyrene plastic coated with 0.1% gelatin. Following rapid thawing and slow dilution in the final culture medium, the initial seeding density should be approximately 20,000 cells/cm2. Cells should be maintained at 37°C in a humidified incubator preferably with 5% CO₂ and 5% oxygen with media change at least twice per week. Upon reaching confluence they should be split 1:3 for routine maintenance. Note: confluence for more than two days may lead to terminal differentiation.

Available Differentiation Kits:

Differentiation Kits: Cat No. ES-K10

Population Doubling Time: Approximately 50 hours.

Population Doubling: Progenitor line RASMO12 is sold at passage 12 (original clonal isolate in confluent 1.9 cm2 well being P1), which corresponds to approximately 15 doublings since the first 1.9 cm2 well and approximately 31.5 doublings since its original clonal plating. The line displays a finite lifespan in vitro, senescing at approximate­ly 70 PD. When used properly this product should scale for a minimum of 10 population doublings.

Quality Control:

Sterility: The line is negative for HIV (1, 2), HBV, HCV, EBV, bacteria, mycoplasma, and fungal contamination.

Thaw Test Result: >50% viability, >25% attachment, growth to confluence, and maintenance of original morpho­logic appearance.

Certificates of Analysis: Available on request.

Restrictions: These cells are provided for research purposes only. They are not for human use, and may not be used for commercial purposes. The user is responsible for proper handling upon receipt.

Technical Assistance: Please contact a representative at ES Cell International (technicalsupport@embryome.com or (510) 521-3390) for technical assistance.

Pricing:

Cat Number Description List Price

ES-154 RASMO12 Embryonic Progenitor Cell Line $1,350.00

ES-M181 ESpan Cell Culture Medium Kit (with supplements) 129.00

ES-K10 Differentiation Kit 295.00

References:

Lusis M, Li J, Ineson J, Christensen ME, Rice A, Little MH. 2010. Isolation of clonogenic, long-term self renewing embryonic renal stem cells. Stem Cell Res. 2010 Mar 27. [Epub ahead of print].

Rasola A, Galietta LJ, Barone V, Romeo G, Bagnasco S. 1995. Molecular cloning and functional characterization of a GABA/betaine transporter from human kidney. FEBS Lett. Oct 16;373(3):229-33.

West, M.D., Sargent, R.G., Long, J., Brown, C., Chu, J-S., Kessler, S., Derugin, N., Sampathkumar, J., Burrows, C., Vaziri, H., Williams, R., Chapman, K.B., Larocca, D., Loring, J.F., and Murai, J. 2008. The ACTCellerate Initiative: large-scale combinatorial cloning of novel human embryonic stem cell derivatives. Reg. Med. 3(3): 287-308.