Steroid-mediated differentiation of neural/neuronal cells from epithelial ovarian precursors in vitro

Abstract

We reported earlier that occasional neurons evolve in human cultures of pluripotent ovarian epithelial stem cells. In subsequent experiments, frequent transdifferentiation into neural stem cells (NSC) and differentiating neurons was observed in human ovarian epithelial stem cells and porcine granulosa cells after exposure to certain combinations of sex steroids. Testosterone (TS), progesterone (PG) or estradiol (E2) alone do not increase the emergence of neurons. However, a mixture of TS + PG after E2 pretreatment converted a majority of ovarian epithelial stem cells or porcine granulosa cells into NSC and differentiating neuronal cells within one to three hours. Cultured neurons manifested an interconnectivity resembling primitive neuronal pathways in culture. These converted cells expressed the cell markers SSEA-1, SSEA-4, NCAM and Thy-1 glycoconjugates of NSC and neurons, and differentiating cells showed characteristic neuronal morphology. Emergence of NSC and neuronal cells was associated with significant cellular depletion of L-glutamic acid (glutamate), which serves as the major excitatory neurotransmitter in the vertebrate CNS and its fast removal is essential for preventing glutamate excitotoxicity. These observations suggest that certain sequential systemic treatment with common sex steroids and their mixture might be effective in the treatment or prevention of degenerative CNS disorders. The ovarian stem cell cultures readily obtainable from human ovaries regardless of the woman's age have the potential to produce NSC for autologous regenerative treatment of neurologic diseases in aging women. Finally, the proper combination of sex steroids could possibly be employed for transdifferentiation of adult bone marrow stem cells or mobilized peripheral blood cells into autologous NSC and stimulate their neuronal differentiation after homing in the CNS.

Sex steroid-mediated reprogramming of vascular smooth muscle cells to stem cells and neurons: Possible utilization of sex steroid combinations for regenerative treatment without utilization of in vitro developed stem cells

“…As a continuation of our recent study dealing with ovarian epithelial cells 2 we tested the effect of Neurobasal/B27 neuron culture medium used for growth and long-term viability of hippocampal neurons (B27 medium) 8 on cultured OSE cells. Among other components, this medium contains undisclosed concentrations of progesterone and retinol.…”

“…1 Regarding transdifferentiation into neurons, umbilical cord blood, amniotic epithelial cells, bone marrow stem cells, and mobilized peripheral blood CD133 + cells can be transdifferentiated into neural stem cells (NSC) by fibroblast growth factor-1, 12-Otetradecanoylphorbol-13-acetate (protein kinase C activator), isobutylmethylxanthine (a Sex steroid-mediated reprogramming of vascular smooth muscle cells to stem cells and neurons non-specific inhibitor of phosphodiesterases), and forskolin (protein kinase A activator), or all-trans-retinoic acid and 2-merkaptoethanol (reviewed in ref. 2). These substances are not suitable for treatment in vivo, however.…”

“…2 In present study we have chosen primary cultures of human vascular smooth muscle cells (SMC), a non-epithelial mesenchymal cells known to express sex steroid receptors [4][5][6] in order to test them as a control cell type regarding their morphology and expression of NSC and neuronal markers. Unexpectedly, however, the vascular SMC subjected to sex steroid combinations also transdifferentiated into NSC and differentiating neuronal cells.…”

Plasticity of granulosa cells: on the crossroad of stemness and transdifferentiation potential

“…Bukovsky et al [9] observed that when porcine ovarian granulosa cells were cultured in vitro, they changed their morphology into fibroepithelial phenotype. Moreover, after exposure of cells to the mixture of testosterone and progesterone after pretreatment with estradiol, a majority of granulosa cells transdifferentiated into neuron-like cells.…”

Ovarian Stem Cell Niche and Follicular Renewal in Mammals

“…Within 10 min (panel B), however, the optically dense organelles are apparent in the extended arm (solid black arrow) within adjacent oocyte cytoplasm (red arrowhead) and distant oocyte regions (red arrow). In reality, normal granulosa cells exhibit fibroblastlike morphology in culture (Bukovsky et al, 2008b), and in vivo the granulosa cells originate from the same precursors as germ cell, that is, the OSC (Bukovsky et al, 1995(Bukovsky et al, , 2004.…”