The clonal human embryonic progenitor cell line EN1 was derived from the registered parental hES cell line H9 (WA09) as described (West et al, 2008). It expresses the distal HOX gene expression HOXA5, B8, C8. Upon differentiation with kit ES-K5, the line expresses the channel SLC7A10 expressed in the choroid plexus, and is therefore useful in the study of the blood brain barrier (Helboe et al, 2003), the cannabinoid receptor 1 (brain) (CNR1), transcript variant 1, mRNA, useful in drug screening, and primary ciliary dyskinesia protein 1 (PCDP1), RASGRP3, and the cytochrome P450 gene CYP3A7. In response to dexamethasone, the line induces PNMT regulating epinephrine synthesis in the adrenal medulla. Unlike the cell line U31, the line EN1 does not express TH.

Positive mRNA Markers: GJB2, KCNN2, CNR1, PNMT, PCDP1, RASGRP3, CYP3A7

Product Information:

Derivation: Progenitor line EN1 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-M40

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 Kit: Cat No. ES-K5

Population Doubling Time: Approximately 50 hours.

Population Doubling: Progenitor line EN1 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-173 EN1 Embryonic Progenitor Cell Line $1,350.00

ES-M40 ESpan Cell Culture Medium Kit (with supplements) 129.00

ES-K5 Differentiation Kit 295.00

References:

Helboe L, Egebjerg J, Møller M, Thomsen C. 2003. Distribution and pharmacology of alanine-serine-cysteine trans­porter 1 (asc-1) in rodent brain. Eur J Neurosci. Oct;18(8):2227-38.

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.