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Hormonally active nontransformed human ovarian cell culture from oophorectomy specimens: methods of development and initial characterization

Seto-Young, Donna; Leonardi, Olga; Park, Alice; Holcomb, Kevin; Salehi, Marzieh; Chang, Peter; Yih, Melissa; Rosenwaks, Zev; Poretsky, Leonid
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
PMID: 16260896
ISSN: 0301-0163
CID: 110201

Beta-cell apoptosis in the pathogenesis of human type 2 diabetes mellitus

Leonardi, Olga; Mints, Gregory; Hussain, Mehboob A
PMID: 12887285
ISSN: 0804-4643
CID: 39126

Brain-derived neurotrophic factor regulates maturation of the DARPP-32 phenotype in striatal medium spiny neurons: studies in vivo and in vitro

Ivkovic S; Polonskaia O; Farinas I; Ehrlich ME
The medium spiny neuron is the predominant striatal neuronal subtype. The striatum, a participant in motor and cognitive functions, is a site of pathophysiology in prevalent neuropsychiatric diseases and is the target of many currently utilized pharmacologic agents. DARPP-32, a dopamine and cyclic AMP-regulated phosphoprotein, is a widely-used marker of mature striatal medium-sized neurons, but the molecules regulating DARPP-32 transcription have not been identified. We show that a null mutation in the mouse brain-derived neurotrophic factor gene leads to decreased DARPP-32 immunoreactivity in striatal medium spiny neurons at birth and postnatal day 10. Striatal DARPP-32 messenger RNA and protein are decreased relative to wild-type littermate controls. In densely plated (1 x 10(6) cells/cm2) primary cultures derived from the ganglionic eminences, addition of brain-derived neurotrophic factor (100 ng/ml) to defined media results in a greater than 3-fold increase in the number of DARPP-32-immunopositive cells after 12 h and greater than 4-fold (P<0.005) after 24 h. The increase in DARPP-32-immunopositivity is abolished by the addition of 2 microg/ml actinomycin D without a significant effect on cell viability. These data suggest that brain-derived neurotrophic factor directly or indirectly regulates DARPP-32 transcription in medium spiny neurons. This is the first demonstration of transcriptional regulation of DARPP-32, and the first evidence of a forebrain abnormality in a newborn neurotrophin 'knockout' mouse
PMID: 9200733
ISSN: 0306-4522
CID: 56955

Doses of peptide growth factors which promote terminal differentiation of E13 to PND1 striatal progenitor cells do not promote long-term survival [Meeting Abstract]

Ivkovic, S.; Polonskaia, O.; Ehrlich, M. E.
BIOSIS:PREV199799769345
ISSN: 0190-5295
CID: 92594

Brain derived neurotrophic factor (BDNF) regulates striatal DARPP-32 and ARPP-21 in vivo and in vitro [Meeting Abstract]

Ivkovic, S.; Polonskaia, O.; Ehrlich, M. E.
BIOSIS:PREV199699218200
ISSN: 0190-5295
CID: 92595