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TM4
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JP Mather
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Biosafety Level:
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1
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frozen
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Medium & Serum:
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See Propagation
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adherent
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Mus musculus (mouse)
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epithelial
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Organ: testis
Disease: normal
Cell Type: Sertoli cell;
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retinol binding protein
tissue plasminogen activator
transferrin
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transfection host (Roche FuGENE® Transfection Reagents)
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follicle stimulating hormone (FSH), expressed
androgen receptor, expressed
progesterone receptor, expressed
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No
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H-Y antigen; Mus musculus, expressed
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11 to 13 days
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male
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ATCC complete growth medium: A 1:1 mixture of Ham'S F12 medium and Dulbecco's modified Eagle's medium with 1.2 g/L sodium bicarbonate and 15 mM HEPES, 92.5%; horse serum, 5%; fetal bovine serum, 2.5%
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
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Protocol:
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Remove and discard culture medium.
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Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
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Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37C to facilitate dispersal.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell supension to new culture vessels.
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Incubate cultures at 37C.
Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:50 is recommended
Medium Renewal: Every 3 to 4 days
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Freeze medium: Culture medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
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Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006
recommended serum:ATCC 30-2020
recommended serum:ATCC 30-2040
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1158: Mather JP. Establishment and characterization of two distinct mouse testicular epithelial cell lines. Biol. Reprod. 23: 243-252, 1980. PubMed: 6774781
1159: Mather JP, et al. Culture of testicular cells in hormone-supplemented serum-free medium. Ann. N.Y. Acad. Sci. 383: 44-68, 1982. PubMed: 7046561
1184: Carson DD, et al. Synthesis and secretion of a novel binding protein for retinol by a cell line derived from Sertoli cells. J. Biol. Chem. 259: 3117-3123, 1984. PubMed: 6538197
26150: Mather JP, Phillips DM. Establishment of a peritubular myoid-like cell line and interactions between established testicular cell lines in culture. J. Ultrastruct. Res. 87: 263-274, 1984. PubMed: 6544874
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