Organ: testis Disease: normal Cell Type: Sertoli cell;
Cellular Products:
retinol binding protein
tissue plasminogen activator
transferrin
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The TM4 cell line is reported to respond to FSH with an increase in cAMP production, but to not respond to luteinizing hormone (LH). The FSH responsiveness is much reduced compared to primary sertoli cell cultures. Constitutive plasminogen activator production is reported to be low, but is stimulated by FSH and, to a greater extent, by retinoic acid.
Tested and found negative for ectromelia virus (mousepox).
Propagation:
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
Subculturing:
Protocol:
Remove and discard culture medium.
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.
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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell supension to new culture vessels.
Incubate cultures at 37C.
Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:50 is recommended Medium Renewal: Every 3 to 4 days
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
References:
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
