Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.
Bottom Line: Transmission electron microscopy revealed the presence of brush border microvilli and tight intercellular contacts.RNA sequencing showed tubular epithelial transcript abundance and revealed the upregulation of components of the EGFR pathway.Thus, reprogrammed BMSCs are a promising cell resource for future cell therapy.
Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: email@example.com.Show MeSH
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Mentions: To explore the renal identity of BMSCs treated with HK2 cell extracts, we studied candidate proximal tubular epithelial cell markers using immunofluorescence microscopy and flow cytometry (fluorescence-activated cell sorting [FACS]) analysis. BMSCs consistently expressed endoglin (ENG; also known as CD105), characteristic of the mesenchymal lineage, and were devoid of the epithelial markers E-cadherin (CDH1) and aquaporin-1 (AQP1) (Figures S1A–S1D). After 8 days, BMSCs treated with HK2 cell extracts weakly expressed CDH1, TJP1 (tight junction protein 1; also known as zona occludens-1), and AQP1 in combination with BMSC markers (Figures 2A–2D). Moreover, after 25 days, BMSCs treated with HK2 cell extracts displayed robust expression of the epithelial markers but substantially reduced expression of ENG (Figures 2E and 2H), which is consistent with the expression profile of HK2 cells (Figures S1E–S1H). Notably, BMSCs that were not converted to cobblestone islands maintained the expression of mesenchymal markers (Figures 2E and 2H, area defined by the dotted line). BMSCs grown in HK2 cell medium, used as control, did not express epithelial markers and retained the expression of ENG (Figures S1I–S1L).
Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: firstname.lastname@example.org.