The clonal human embryonic progenitor cell line U31 was derived from the registered parental hES cell line H9 (WA09) as described (West et al, 2008). It expresses markers of dopa­mine metabolism and sympathoblastic neural crest including the neurotrophin receptor NTRK, TH, CRABP1, KIT, and no HOX gene expression. Upon differentiation with kit ES-K43, the line expresses the channels SLC13A4, the neurotransmitter transporters SLC6A1 and SLC6A12, expressed in the meninges, and is therefore useful in the study of the blood brain barrier and drug discovery (Thoeringer et al, 2009) as well as high levels of PPP1R1B (DARPP32), a downstream target of the dopamine receptor. When differentiated in the presence of dexamethasone, the line expresses PNMT a final step in the synthesis of epinephrine.

Positive mRNA Markers: NTRK, TH, CRABP1, KIT, DARPP32, SLC13A4, PNMT

Product Information:

Derivation: Progenitor line U31 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-NC100

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-K43

Population Doubling Time: Approximately 50 hours.

Population Doubling: Progenitor line U31 is sold at passage 13 (original clonal isolate in confluent 1.9 cm2 well being P1), which corresponds to approximately 16.5 doublings since the first 1.9 cm2 well and approximately 33 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 bacteria, 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-236 U31 Embryonic Progenitor Cell Line $1,350.00

ES-NC100 ESpan Cell Culture Medium Kit (with supplements) 129.00

ES-K43 Differentiation Kit 295.00

References:

Thoeringer CK, Ripke S, Unschuld PG, Lucae S, Ising M, Bettecken T, Uhr M, Keck ME, Mueller-Myhsok B, Holsboer F, Binder EB, Erhardt A. 2009. The GABA transporter 1 (SLC6A1): a novel candidate gene for anxiety disorders. J Neural Transm. Jun;116(6):649-57. Epub 2008 Jul 8.

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.