Effect of cell purity,cell concentration,and incubation conditions on rat testis leydig cell steroidogenesis |
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Authors: | D R E Abayasekara L O Kurlak A M Band M H F Sullivan B A Cooke |
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Institution: | (1) Present address: MRC/AFRC Comparative Physiology Group, Institute of Zoology, Zoological Society of London, Regent’s Park, NW1 4RY London, United Kingdom;(2) Department of Biochemistry, Royal Free Hospital School of Medicine, Rowland Hill Street, NW3 2PF London, UK |
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Abstract: | Summary This study examines the effects of cell purity and incubation conditions on testosterone production by rat testis Leydig cells
in short-term primary culture. Both basal and luteinizing hormone (LH)-stimulated testosterone production were affected by
the purity of the cell preparation, i.e. as the purity of the cell preparation was increased the amount of testosterone produced
per Leydig cell was also found to increase. The stimulation ratio of testosterone production, calculated as the secretion
of testosterone in the presence of LH (100 ng/ml) divided by the basal secretion of testosterone, increased with the increase
in plating density (20 000 to 200 000 cells per well). This pattern of change was independent of the vessel and volume of
incubation. In terms of the absolute amount of testosterone produced, increasing the plating density led to a decrease in
the amount of steroid produced both basally and in response to LH. Composition of the incubation medium also had an effect
on testosterone production; phenol red and sodium bicarbonate exerted negative effects. At all temperatures studied (4°, 24°,
34°, and 37° C), LH increased testosterone production and the degree of stimulation increased with temperature. We conclude
that cell purity and incubation conditions markedly affect rat Leydig cell steroidogenesis in vitro. Furthermore, the manner
in which the results are presented can affect their interpretation. |
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Keywords: | Leydig cell cell purity cell concentration luteinizing hormone steroidogenesis testosterone |
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