Immortalized Human Telomerized Endometrial Stromal Cells (SHT290)

Endometrial stromal cells derived from biopsies were immortalized with pDSWK-8 which encodes a version of the retroviral pBABEpuro-TERT to establish Immortalized Human Telomerized Endometrial Stromal Cells (SHT290). SHT290 cells express decidualization markers (prolactin and plasminofen inhibitor type-1). The endometrial stromal cells may be differentiated to support hormonal effects on co-cultured endometrial epithelial cells, estrogen and progesterone must be cycled in the medium to simulate hormonal variations during the menstrual cycle.

Immortalization Method: Immortalized with retroviral transduction containing hTERT

BioSafety Level: II

Organism: Human (H. sapiens)

Species: Human

Source Organs: Reproductive

Organ Type: Reproductive

Growth Properties: Adherent

Morphology: Uniform bipolar, fibroblast-like shape

Population Doubling: 4 - 6 days

Markers: Puromycin, prolactin, plasminofen inhibitor type-1, vimentin, cytokeratin 13, cytokeratin 18

Donor Age: Not disclosed

Donor Gender: Female

Propagation: Note: Initial seeding and subculturing must done at high density or the cells will cease to grow. Cells should be grown in the absence of puromycin. The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add charcoal-stripped fetal bovine serum to a final concentration of 2%, 1X (2mM)L-glutamine (G275), and, Penicillin/Streptomycin Solution (G255).Change media every 2-3 days.Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise. To adapt for co-culture with epithelial cells: The base medium is a 1:1 mixture of Ham’s F12 (Gibco, Invitrogen Corp., Carlsbad, CA) and M199 basic medium (Sigma, St. Louis, MO) supplemented with 5% bovine calf serum (BCS; Hyclone, Logan, UT), 0.1% Mitoplus (BD Biosciences, Becton Dickinson, Franklin Lakes, NJ), 2 µg/ml of insulin (Sigma) and antibiotic/antimycotic solution diluted to yield 100 units/ml penicillin G sodium, 100 mg/ml streptomycin sulfate and 250 ng/ml amphotericin B (ABAM; Gibco). Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise. Decidualization procedure: The base medium for this procedure is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add fetal bovine serum to a final concentration of 2%, 1X (2mM)L-glutamine (G275), 20 ng/ml human EGF (Z100135)and, Penicillin/Streptomycin Solution (G255).Change media every 2-3 days.Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Epifluorescence localization of cytoskeletal proteins conducted with mouse monoclonal antibodies against vimentin, cytokeratin 13, and cytokeratin 18

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C

Reference: Barbier, C. S., Becker, K. A., Troester, M. A., & Kaufman, D. G. (2005). Expression of exogenous human telomerase in cultures of endometrial stromal cells does not alter their hormone responsiveness. Biology of reproduction, 73(1), 106–114. https://doi.org/10.1095/biolreprod.104.035063Barbier, C., Kloosterboer, H. J., & Kaufman, D. G. (2008). Effects of tibolone metabolites on human endometrial cell lines in co-culture. Reproductive sciences (Thousand Oaks, Calif.), 15(1), 75–82. https://doi.org/10.1177/19337191073097198