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Immortalized Rat Steroidogenic Granulosa Cells stably expressing LH/CG Receptor

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Immortalized Rat Steroidogenic Granulosa Cells stably expressing LH/CG Receptor

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Immortalized Rat Steroidogenic Granulosa Cells stably expressing LH/CG Receptor

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	Print Version
BioSafety Level	II
Organism	Immature female rat
Source Organ	Ovary
Growth Properties	Adherent
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
Immortalization Method	Serial passaging and calcium phosphate mediated cotransfection by SV40 DNA and Ha-ras oncogene
Description	Lutropin/choriogonadotropin (LH/CG) receptor, a receptor uniquely expressed in ovarian granulosa and testicular sertoli cells, play an important role in the regulation of oogenesis, spermatogenesis and steroid hormone production. Unfortunately the LH/CG receptor expression is often lost within 1-2 days of in vitro culture of primary cells. The Immortalized Rat Steroidogenic Granulosa Cells stably expressing LH/CG Receptor is responsive to luteinizing hormone (LH) or choriogonadtropin (hCG) stimulation and produces progesterone and 20α-OH-P as a result at levels comparable to primary cells. This cell line is useful in studying molecular mechanisms associated with gonadotropin stimulated steroidogenesis. Together with the Immortalized Rat Steroidogenic Granulosa Cells expressing FSH Receptor (Cat. No T0605), researchers can compare the LH and FSH signaling in homologues cell systems.
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow IV medium available from ABM (TM004). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 37.0°C. To stimulate progesterone production, add 6 nM hCG or 1.2 nM LH, and 2 μM forskolin to the complete culture medium for at least 24 hours.
Quality Control	1) Phenotypic response to hCG stimulation was characterized by shape changes (cells rounding up as a result of intracellular cAMP accumulation); 2) LH/CG receptor expression was measured by radiolabelled 125I-hCG binding assay- the cells express 19,000 FSH receptors/cell with a Kd of 180 pM, similar to the native receptor; 3) Production of progesterone and 20α-OH-P was determined by radioimmunoassay (RIA); 4) Production of cAMP was measured by a protein binding method
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.