OPTIMIZING TESTOSTERONE PRODUCTION BY PURIFIED ADULT-RAT LEYDIG-CELLS INVITRO
OPTIMIZING TESTOSTERONE PRODUCTION BY PURIFIED ADULT-RAT LEYDIG-CELLS INVITRO
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DOI：
10.1007/bf02629089
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发表时间：
1988-06-01
期刊：
影响因子：
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通讯作者：
EWING, LL
中科院分区：
文献类型：
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作者：
KLINEFELTER, GR;EWING, LL
我们试图建立在原代培养中增加完全分化的成年大鼠睾丸间质细胞产生睾酮的条件。新鲜分离的高纯度成年大鼠睾丸间质细胞悬液，在最大刺激浓度为绵羊黄体生成素(LH)的情况下，在1.5ml微离聚管中培养3h，产生睾酮83 ng/106Leydig细胞·h−1。不幸的是，只能附着在塑料培养皿上的成年大鼠睾丸间质细胞变平了，睾丸激素的产生迅速减少。含Cytodex 3小球的改良Eagles‘s Ham’s F12(1：1；VOL/VOL)培养液中的间质细胞在含有胎牛血清的培养液中预平衡后仍能存活，但在类似刺激条件下孵育24 h，睾丸细胞仅产生30 ng睾酮/10~6Leydig细胞·h−1。同样培养的间质细胞在黄体生成素的刺激下，与1 mg/ml的牛脂蛋白共同孵育时，对牛脂蛋白(1.222 g/ml)产生105ng睾酮/106间质细胞·h−1的反应。因此，在原代培养中，脂蛋白可维持纯化的成年大鼠间质细胞的类固醇生成能力24小时。
We sought to establish conditions that increased the duration of testosterone production by fully differentiated adult rat Leydig cells in primary culture. A freshly isolated suspension of highly purified adult rat Leydig cells produced 83 ng testosterone/106Leydig cells·h−1when incubated in Medium 199 in a 1.5 ml microfuge tube with shaking for 3 h with a maximally stimulating concentration of ovine luteinizing hormone (LH). Unfortunately, adult rat Leydig cells that were allowed to attach only to a plastic culture dish flattened out, and testosterone production diminished rapidly. Leydig cells in Dulbecco's modified Eagles' medium-Ham's F12 (1∶1; vol/vol) containing Cytodex 3 beads pre-equilibrated in culture medium containing fetal bovine serum attached to the beads and remained viable, but produced only 30 ng testosterone/106Leydig cells·h−1when incubated for 24 h with similar stimulation. Leydig cells similarly cultured and maximally stimulated with LH, responded to bovine lipoproteins (<1.222 g/ml) producing 105 ng of testosterone/106Leydig cells·h−1when incubated with 1 mg/ml bovine lipoprotein. Therefore, lipoproteins maintain the steroidogenic capacity of purified adult rat Leydig cells in primary culture for 24 h.