Immortalized Rat Steroidogenic Granulosa Cells stably expressing FSH Receptor (GFSHR-17)

CAT.NOUNITPRICE
T06051x106 cells / 1.0 ml
$0.00

Specifications


DescriptionFollicle stimulating hormone (FSH) 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 FSH receptor expression is often lost within 1-2 days of in vitro culture of primary cells.

The Immortalized Rat Steroidogenic Granulosa Cells stably expressing FSH Receptor is responsive to FSH stimulation and produces progesterone as a result at levels comparable to primary cells. This cell line is useful in studying FSH-FSH receptor interactions, as well as how FSH receptor regulates the development and maturation of ovarian follicle. Together with the Immortalized Rat Steroidogenic Granulosa Cells expressing LH Receptor (Cat. No T0606), researchers can compare the LH and FSH signaling in homologues cell systems.
SKUT0605
SpeciesRat (R. norvegicus)
Tissue/Organ/Organ SystemReproductive
Cell MorphologyMulti-Polar
ApplicationsFor Research Use Only
Seeding Density10,000 - 20,000 cells/cm2
Population Doubling Time35 - 45 hours
Immortalization MethodSerial passaging and calcium phosphate mediated cotransfection by SV40 DNA and Ha-ras oncogene
Cell TypeImmortalized Cells
Growth PropertiesAdherent
Propagation Requirements
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.
The base medium for this cell line is Prigrow IV medium available at 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 Solution (G255) to a final concentration of 1%
Carbon dioxide (CO2): 5%, Temperature: 37.0
Subculture Protocol0
Unit quantity1x106 cells / 1.0 ml
QC1) Phenotypic response to FSH was characterized by shape changes (cells rounding up); 2) FSH receptor expression was measured by radiolabelled 125I-oFSH binding assay- the cells express 27,000 FSH receptors/cell with a Kd of 100-115 pM, similar to the native receptor; 3) Production of progesterone was determined by radioimmunoassay (RIA); 4) Production of cAMP was measured by a protein binding method
Disclaimer

1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item.

2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].

3. 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.

4. We 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).

5. 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.

6. 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.

7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."

FAQs


I want to make sure these cells express my gene of interest before I decide to buy the cell line. Can you provide a sample so this can be tested?
We do not carry out downstream characterization or gene expression profiling of our cell lines. To facilitate your preliminary experiments we can provide an RNA extraction (0.5ug total RNA) or cell lysate (100ug/100ul provided in 62.5mM Tris‐HCl, 2% SDS, 10% Glycerol, 50mM DTT, 0.01% w/v Bromophenol Blue) for any of our immortalized cell lines for a small fee. Please inquire directly for more information. The lead time will be around 2 weeks from the time of placing an order (if the item is in stock).
How often do I need to change the media?
The media should be changed every 2-3 days.
Why do these cells need bio safety level II?
In order to be more cautious, we follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site if required.
Do you sell ECM coated T75 flasks?
Yes we can provide a coating service. Please inquire with [email protected]
What can I coat a larger dish to subculture?
We also offer applied extracellular matrix (collagen type I) in liquid form, for the coating of larger flasks and other required plasticware: http://www.abmgood.com/Applied-Cell-Extracellular-Matrix-G422.html
How long can I store frozen vials for?
Cells that are properly frozen using an effective cryoprotective agent can be stored in liquid nitrogen indefinitely without affecting their recovery.
Should the cap of the flask be changed before starting the cell culturing step?
No, there is no need in sterile biosafety cabinets unless it has contacted any non-sterile condition (e.g. touching the contaminated tip, etc.).
What is the recommended storage temperature?
In general, if you received: Live cells: acclimatize for 3-4 hrs at at the recommended conditions stated for the cell line under the propagation section, and then change media afterwards. Frozen cells: Immediately place cells in liquid nitrogen; -180C.
How is cell density crucial for drug selection?
If antibiotic selection is applicable to the target cells, we suggest getting rid of all the background cells so that the cell density is kept lower (even 20-30%). However, once the clones are selected by clonal dilution, we don't need the drug to still be present. If needed, the cell density should be towards the higher end since cells are already selected. Any primary cells still present will be depleted as a result of senescence and the cell population that remains will be resistant to the specific antibiotic.
My cells are not detaching, what method do you recommend to trypsinize the cells?
1. Incubate the coated plate containing trypsin solution at recommended temperature indicated in the propagation section for 3-5 min till the cells round up, monitoring from time to time under microscope. 2. Diluting G422 (1:1) with PBS and coating for lesser time. Sometimes the collagen content in G422 is higher and thus make stronger bonding with cells. 3. You can try reducing the incubation time as well for coating the plate to make a thinner layer.
Why is it important to determine the optimal seeding density?
The seeding density we recommend is for when cells are plated to a new vessel. The optimal seeding density should allow cells to attach to the surface and have room to proliferate. If you seed too little, cells may not attach well to the surface (for adherent cells). Seeding density is important as many cells (adherent or suspension cells) need to be in close proximity for better growth. Cell-cell interactions allow cells to communicate with each other in response to changes in their microenvironment. This ability to send and receive signals is essential for the survival of the cell. In other cases, if the seeding density is too low, cells may attach but a retardation in cell growth is observed. If you seed too high, the cells will attach but there is insufficient room for further proliferation and they will stop replicating.
References


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  • Keren-Tal, I et al. "Establishment of Steroidogenic Granulosa Cell Lines Expressing Follicle Stimulating Hormone Receptors" Molecular and Cellular Endocrinology 95(1-2):R1-R10 (1993). PubMed: 8243796.
  • Shukovski, L et al. "Regulation of Follistatin Messenger Ribonucleic Acid in Steroidogenic Rat Granulosa Cell Lines" Endocrinology 136(70):2889-95 (1996). PubMed: 7789314.
  • Keren-Tal, I et al. "Activation of FSH-Responsive Adenylate Cyclase by Staurosporine: Role for Protein Phosphorylation in Gonadotropin Receptor Desensitization" Mol Cell Endocrinol 116(1):39-48 (1996). PubMed: 8822263.
  • Schiffer, Z et al. "Fourier Analysis of Differential Light Scattering for the Quantitation of FSH Response Associated with Structural Changes in Immortalized Granulosa Cells" Mol Cell Endocrinol 118(1-2):145-5 (1996). PubMed: 8735600.
  • Selvaraj, N et al. "Partial Sequencing of the Rat Steroidogenic Acute Regulatory Protein Message from Immortalized Granulosa Cells: Regulation by Gonadotropins and Isoproterenol" Mol Cell Endocrinol 123(2):171-7 (1996). PubMed: 8961254.
  • Selvaraj, N et al. "Modulation of FSH Receptor Phosphorylation Correlates with Hormone-Induced Coupling to the Adenylate Cyclase System" Endocrine 6(2):179-85 (1997). PubMed: 9225133.
  • Grosse, J et al. "Synaptosome-Associated Protein of 25 Kilodaltons in Oocytes and Steroid-Producing Cells of Rat and Human Ovary: Molecular Analysis and Regulation by Gonadotropins" Biol Reprod 63(2):643-50 (2000). PubMed: 10906076.
  • Sommersberg, B et al. "Gap Junction Communication and Connexin 43 Gene Expression in a Rat Granulosa Cell Line: Regulation by Follicle-Stimulating Hormone" Biol Reprod 63(6):1661-8 (2000). PubMed: 11090433.
  • Ngezahayo, A et al. "Regulation of Ion Fluxes, Cell Volume and Gap Junctional Coupling by cGMP in GFSHR-17 Granulosa Cells" J Membr Biol 194(3):165-76 (2003). PubMed: 14502429.
  • Ngezahayo, A et al. "Gap Junction Coupling and Apoptosis in GFSHR-17 Granulosa Cells" J Membr Biol 204(3):137-44 (2005). PubMed: 16245036.
  • Mayerhofer, A et al. "FSH Regulates Acetycholine Production by Ovarian Granulosa Cells." Reprod Biol Endocrinol 17(4):37 (2006). PubMed: 16846505.
  • Schlie, S et al. "Three-Dimensional Cell Growth on Structures Fabricated from ORMOCER by Two-Photon Polymerization Technique" J Biomater Appl 22(3):275-87 (2007). PubMed: 17494962.
  • Ovsianikov, A et al. "Two-Photon Polymerization Technique for Microfabrication of CAD-Designed 3D Scaffolds from Commercially Available Photosensitive Materials" J Tissue Eng Regen Med 1(6):443-9 (2007). PubMed: 18265416.
  • Baumgart, J et al. "Quantified Femtosecond Laser Based Opto-Perforation of Living GFSHR-17 and MTH53 a cells" Opt Express 16(5):3021-31 (2008). PubMed: 18542388 .
  • Bintig, W et al. "Purinergic Signalling in Rat GFSHR-17 Granulosa Cells: An in vitro Model of Granulosa Cells in Maturing Follicles" J Bioenerg Biomembr 41(1):85-94 (2009). PubMed: 19191015.