Hormonally active nontransformed human ovarian cell culture from oophorectomy specimens: methods of development and initial characterization.
We repeatedly established a nontransformed steroidogenically active human ovarian cell culture derived from oophorectomy specimens. The cells maintained steroidogenic activity for 3-5 passages (6-8 weeks) and responded to stimulation by insulin and gonadotropin. With pregnenolone as substrate, LH stimulated progesterone production up to 124% and FSH up to 121%. Insulin alone stimulated progesterone production up to 135%, in the presence of LH up to 191%, and in the presence of FSH up to 170%. With dehydroisoandrosterone (DHA) as substrate, insulin alone stimulated testosterone production up to 117%, and in the presence of LH (but not FSH) up to 125%. With androstenedione as substrate, insulin alone stimulated estradiol production up to 133%, FSH alone up to 188%, and LH with insulin up to 217%. With progesterone as substrate and in the presence of LH (but not FSH), 17-alpha-hydroxylase activity was stimulated up to 131%. With DHA as substrate and in the presence of LH, 3-beta-hydroxysteroid dehydrogenase (3-beta-HSD) activity was stimulated up to 139%. With androstenedione as substrate, insulin alone stimulated aromatase activity up to 202%, LH up to 208%, and FSH up to 251%. Under the same conditions, in the presence of LH and insulin, aromatase activity was stimulated up to 342%, and in the presence of FSH and insulin, up to 318%. With testosterone as substrate, insulin alone stimulated aromatase activity up to 122%. With testosterone as substrate, in the presence of LH and insulin, aromatase activity was stimulated up to 136%, and in the presence of FSH and insulin, up to 156%. Immunocytochemistry studies directly confirmed presence of aromatase and 3-beta-HSD in these cultured cells. We conclude that a steroidogenically active nontransformed long-term human ovarian cell culture can be repeatedly established from oophorectomy specimens, providing uninterrupted supply of cultured human ovarian cells for a variety of studies of ovarian physiology.
Seto-Young
Donna
D
Division of Endocrinology, Department of Medicine, Beth Israel Medical Center and Albert Einstein College of Medicine, NY 10003, USA.
Leonardi
Olga
O
Park
Alice
A
Holcomb
Kevin
K
Salehi
Marzieh
M
Chang
Peter
P
Yih
Melissa
M
Rosenwaks
Zev
Z
Poretsky
Leonid
L
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000328:Adult; D000735:Androstenedione / Q000378:metabolism; D001141:Aromatase / Q000378:metabolism; D002478:Cells, Cultured; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000494:pharmacology; D008875:Middle Aged; D010052:Ovariectomy; D010053:Ovary / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011284:Pregnenolone / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism; D013739:Testosterone / Q000096:biosynthesis
D007328:Insulin; D013739:Testosterone; D000735:Androstenedione; D011374:Progesterone; D004958:Estradiol; D011284:Pregnenolone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D015096:3-Hydroxysteroid Dehydrogenases; D001141:Aromatase; D013254:Steroid 17-alpha-Hydroxylase