Organ: testis Disease: normal Cell Type: Leydig cell;
Cellular Products:
prostaglandin F2a
Permits/Forms:
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The TM3 cell line responds to LH with an increase in cAMP production, but does not respond to follicle stimulating hormone (FSH). The maintenance of responsiveness to LH is dependent upon serum lot. In the presence of LH, the cells are capable of metabolizing cholesterol. 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 2.5mM L-Glutamine, 0.5mM Sodium Pyruvate, 1.2 g/L Sodium Bicarbonate and 15 mM HEPES, 92.5%; horse serum, 5%; fetal bovine serum, 2.5%
Subculturing:
Protocol: Remove medium, add fresh 0.25% trypsin-0.53mM EDTA solution, rinse, remove trypsin and let the culture sit at room temperature for 2 to 4 minutes. Add fresh medium, aspirate and dispense into new flasks. Subc*tion Ratio: A subc*tion ratio of 1:20 to 1:200 is recommended Medium Renewal: Every 3 to 4 days