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Isolation of Human Leydig Cells Which Are Highly Responsive to Human Chorionic Gonadotropin

MRC Reproductive Biology Unit, Centre for Reproductive Biology Edinburgh, EH3 9EW Scotland

Address requests for reprints to: Dr. Barbara Simpson, Medical Research Council Reproductive Biology Unit, Centre for Reproductive Biology, 37 Chalmers Street, Edinburgh, Scotland EH3 9EW.

Leydig cells were purified on discontinuous Percoll gradients after collagenase digestion of human or rat testes. Leydig cells from both species were found in three bands. As determined by positive staining for 3β-hydroxysteroid dehydro-genase, band 1 (lowest density cells) from both species contained only 12–28% Leydig cells. However, while band 3 was the most Leydig cell-enriched fraction in rat cell preparations (70–90% Leydig cells), human band 2 (48–70% Leydig cells) was consistently more Leydig cell enriched than was band 3 (30–56% Leydig cells).

Despite their slightly different fractionation pattern, Leydig cells prepared from five men with prostatic carcinoma were similar to those from the rat, both in terms of the amount of testosterone produced basally per 10⁶ Leydig cells (80–234 ng/20 h) and in terms of the magnitude of their response to hCG (764–1342 ng/10⁶ Leydig cells-20 h; 5- to 17-fold stimulation above basal). Cells prepared from five other men with prostatic carcinoma produced much lower amounts of testosterone, but still had up to a 6-fold response to hCG. Plasma LH, FSH, and testosterone concentrations in the latter group could not distinguished from those in the group whose Leydig cells duced large amounts of testosterone in vitro. Morphologically, the testes from the latter group appeared to contain more darkly staining than lightly staining Leydig cells than did the former group.

Rat Leydig cells responded in a dose-dependent fashion hCG over the range 0.03–0.5 mU/mL, whereas human Leydig cells were 10- to 100-fold less sensitive, responding to hCG the range 0.4–100 mU/mL. The number and affinity of Leydig cell LH (hCG) receptors were assessed from Scatchard analysis of [¹²⁵I]hCG binding. Compared with rat cells, human Leydig cells contained approximately 20% of the number of LH recep tors, while the affinity of the receptors (K_d, 10^–10 M) was similar to that in rats.

In conclusion, a method for the isolation of highly responsive human Leydig cells has been developed. The results so suggest that the function of human Leydig cells may be similar to that of the rat than thought previously. (J Endocrinol Metab 65: 415, 1987)

Received December 30, 1986.

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