|
Print Version |
| BioSafety Level |
II |
| Organism |
Homo sapiens |
| Source Organ |
Ovaries |
| Growth Properties |
Adherent |
| Recommended Seeding Density |
20,000 cells/cm2 |
| Applications |
For Research Use Only |
| Immortalization Method |
Serial passaging and transduction with retroviruses carrying human papilloma virus strain 16 (HPV16) oncogenes E6/E7 |
| Description |
Human ovarian granulosa cells are the sites of estrogen and progesterone production, making them critical in studying steroid biosynthesis, steroid-metabolizing enzymes, particular growth factors, and gonadotropins. The Immortalized Human Granulosa Cells (HGL5) exhibits qualities consistent with primary ovarian granulosa cells; this includes cell retraction in response to protein kinase-A activation, the ability to produce progesterone and estradiol, and the enzyme integral to estradiol production, P450arom. All of these characteristics make the Immortalized Human Granulosa Cell Line an attractive model to study mechanisms relating to steroid biosynthesis and metabolism, in addition to other pathways. |
| Procedure Overview |
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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 in ABM, Cat. No.TM004. To make the complete growth medium, add the following components to the base medium: 2% Cosmic Calf Serum (gibco), 2% Ultroser G (Pall Corporation), 1% ITS plus (Zen-Bio), and 1% Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%.Temperature: 37.0°C.
Low serum media for experiments: The base medium for this cell line is Prigrow IV medium available in ABM, Cat. No.TM004. To make the complete growth medium, add the following components to the base medium: 0.2% Ultroser SF (Pall Corporation), 1% ITS plus (Zen-Bio), and 1% Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%.Temperature: 37.0°C. |
| Subculturing |
1. Dilute Trypsin-EDTA (TM050) to final concentration of 0.05%, and use to trypsinize cells for 3-5 minutes at room temperature
2. Gently tap flask to remove cells, and pipette cells up and down in flask before transferring to sterile conical vial with growth media.
3. Centrifuge cells at 200 x g for 3 minutes.
4. Aspirate supernatant from pellet and resuspend cells with fresh growth medium to plate cells.
Be sure to use T25 ECM-coated flasks(G299) for growth. |
| Preservation |
1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase. |
| Quality Control |
(1) Expression of viral genes quantified by PCR; (2) cAMP, progesterone and estradiol production were measured after induction with forskolin, dbcAMP, follicle-stimulating hormone (FSH), or luteinizing hormone (LH) treatments; (3) p450arom regulation characterized via North blot analysis. |
| 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. |
