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목적 : Direct conversion of somatic cells into lineage-committed cells can simplify the protocols for production of neural stem/progenitor cells. We were to identify the characteristics of lens epithelial cells (LECs) to use as candidate material to reprogramming and to present another single factor, Oct4, also has ability to generate the mouse LECs into the induced neural stem cells (iNSCs).
방법 : Lens epithelial cells were prepared from neonatal mouse lens. Sox2, Gbx2, Mash1, Sox1, Olig2, Brn2, Brn4, and Oct4 were expressed individually in mouse LECs by lentiviral-mediated gene transduction. The primary neurospheres were seeded and allowed to differentiate for 14 days. Molecular characteristics were evaluated in immunochemistry, RT-PCR, and western blot. Electrophysiological Recordings were examined the function of induced cells.
결과 : RT-PCR revealed transcription of SOX2, Pax2, and Nestin, all of which are characteristic makers of NSCs both in LECs and brain-derived NSCs. iNSCs induced by Oct4, express NSC markers and resemble wild-type NSCs in their morphology, self-renewal, and ability to form neurospheres. Cloned iNSCs differentiate into several types of mature neurons, as well as astrocytes and oligodendrocytes, indicating multipotency.
결론 : LECs have similar molecular characteristics with brain-derived NSCs and can be a source of direct reprogramming into self-renewable iNSCs by single factor Oct4. Because there is already endogenous Sox2 in LECs, Oct4 may initiate or trigger the direct conversion into iNSCs.
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