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목적 : Wnt signaling plays an important role in the regulation of self-renewal in stem cells. Here we investigated the effect of modulation of β-catenin, the primary transducer of the Wnt signaling canonical pathway, on the growth and differentiation of the limbal epithelial progenitors according to the culture condition of limbal epithelial cells.
방법 : Limbal explant cultures were made in control medium or medium complemented with IWP2, a Wnt signal blocker, or CHIR99021, an indirect β-catenin activator. We determined outgrowth size from limbal biopsy explants, cell size, protein and mRNA expression of markers for stem/precursor cells for corneal epithelial differentiation, and colony formation efficiency (CFE). Comparative experiments were performed in cultures of freshly isolated limbal epithelial cells grown in low calcium medium.
결과 : Limbal explant outgrowth rate with IWP2 was increased. In the limbal explant cultured cells, the effect of the two Wnt signaling modifiers on ABCG2 expression, function, and CFE were in opposite directions to the effects seen on the isolated limbal epithelial cell cultures. In spite of the divergent effects, in both systems IWP2 decreased and CHIR99021 increased total β-catenin, respectively. The limbal explant culture system with IWP2 increased proliferative potential (p63α), proliferation status (Ki67), and decreased differentiation (Krt12).
결론 : These results suggest that, unlike the case for isolated cell cultures, inhibition of the canonical Wnt pathway has a positive impact on stem/precursor cells in outgrowths from limbal explant cultures.
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