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Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.

Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, Grazia de Simoni M, Remuzzi G, Goligorsky MS, Benigni A - Stem Cell Reports (2015)

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.

View Article: PubMed Central - PubMed

Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: evangelia.papadimou@marionegri.it.

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Human CL17, HK2, and BMSC Integration in Renal Organoids(A) Human CL17, HK2, or BMSCs were mixed with fully dissociated E11.5 mouse embryonic kidneys and grown as explants for 1–5 days. Representative image of CL17-cell integration (stained with Cell Tracker, green) into the condensing metanephric mesenchyme identified by NCAM1 and PAX2 co-staining at day 1 (insert image).(B and C) At day 5, chimeric aggregates of human (immunostained with anti human nuclear antigen, green; arrowhead) and mouse cells (asterisk) lay within laminin-positive membranes (white). Cell nuclei were labeled with DAPI (blue).(E and F) CL17 cells (E) and HK2 cells (F) form elongating tubular structures in the vicinity of WT1-positive glomerular structures.(D and G) BMSCs never formed any renal structure. Images are representative of three independent experiments.
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fig5: Human CL17, HK2, and BMSC Integration in Renal Organoids(A) Human CL17, HK2, or BMSCs were mixed with fully dissociated E11.5 mouse embryonic kidneys and grown as explants for 1–5 days. Representative image of CL17-cell integration (stained with Cell Tracker, green) into the condensing metanephric mesenchyme identified by NCAM1 and PAX2 co-staining at day 1 (insert image).(B and C) At day 5, chimeric aggregates of human (immunostained with anti human nuclear antigen, green; arrowhead) and mouse cells (asterisk) lay within laminin-positive membranes (white). Cell nuclei were labeled with DAPI (blue).(E and F) CL17 cells (E) and HK2 cells (F) form elongating tubular structures in the vicinity of WT1-positive glomerular structures.(D and G) BMSCs never formed any renal structure. Images are representative of three independent experiments.

Mentions: To assess the functional integration into kidney tissue, human cells (CL17, HK2, or BMSCs) were mixed with kidney cells derived from embryonic day 11.5 (E11.5) mice (Unbekandt and Davies, 2010; Xinaris et al., 2012), and grown as explants for 5 days. At day 1, CL17 cells integrated into the condensing metanephric mesenchyme that was identified by co-expression of neural cell adhesion molecule (NCAM) and PAX2 markers (Figure 5A, insert). At day 5, chimeric aggregates of human and mouse cells grew into elongating tubular structures that were surrounded by laminin-positive basement membranes (Figures 5B and 5C). Remarkably, tubular structures containing CL17 or HK2 cells were in close vicinity to glomerular-like structures expressing the early podocyte marker Wilms tumor 1 (WT1) (Figures 5E and 5F). BMSCs neither formed nor contributed to renal structures, indicating non-nephrogenic potential (Figures 5D and 5G).


Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts.

Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, Grazia de Simoni M, Remuzzi G, Goligorsky MS, Benigni A - Stem Cell Reports (2015)

Human CL17, HK2, and BMSC Integration in Renal Organoids(A) Human CL17, HK2, or BMSCs were mixed with fully dissociated E11.5 mouse embryonic kidneys and grown as explants for 1–5 days. Representative image of CL17-cell integration (stained with Cell Tracker, green) into the condensing metanephric mesenchyme identified by NCAM1 and PAX2 co-staining at day 1 (insert image).(B and C) At day 5, chimeric aggregates of human (immunostained with anti human nuclear antigen, green; arrowhead) and mouse cells (asterisk) lay within laminin-positive membranes (white). Cell nuclei were labeled with DAPI (blue).(E and F) CL17 cells (E) and HK2 cells (F) form elongating tubular structures in the vicinity of WT1-positive glomerular structures.(D and G) BMSCs never formed any renal structure. Images are representative of three independent experiments.
© Copyright Policy - CC BY-NC-ND
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fig5: Human CL17, HK2, and BMSC Integration in Renal Organoids(A) Human CL17, HK2, or BMSCs were mixed with fully dissociated E11.5 mouse embryonic kidneys and grown as explants for 1–5 days. Representative image of CL17-cell integration (stained with Cell Tracker, green) into the condensing metanephric mesenchyme identified by NCAM1 and PAX2 co-staining at day 1 (insert image).(B and C) At day 5, chimeric aggregates of human (immunostained with anti human nuclear antigen, green; arrowhead) and mouse cells (asterisk) lay within laminin-positive membranes (white). Cell nuclei were labeled with DAPI (blue).(E and F) CL17 cells (E) and HK2 cells (F) form elongating tubular structures in the vicinity of WT1-positive glomerular structures.(D and G) BMSCs never formed any renal structure. Images are representative of three independent experiments.
Mentions: To assess the functional integration into kidney tissue, human cells (CL17, HK2, or BMSCs) were mixed with kidney cells derived from embryonic day 11.5 (E11.5) mice (Unbekandt and Davies, 2010; Xinaris et al., 2012), and grown as explants for 5 days. At day 1, CL17 cells integrated into the condensing metanephric mesenchyme that was identified by co-expression of neural cell adhesion molecule (NCAM) and PAX2 markers (Figure 5A, insert). At day 5, chimeric aggregates of human and mouse cells grew into elongating tubular structures that were surrounded by laminin-positive basement membranes (Figures 5B and 5C). Remarkably, tubular structures containing CL17 or HK2 cells were in close vicinity to glomerular-like structures expressing the early podocyte marker Wilms tumor 1 (WT1) (Figures 5E and 5F). BMSCs neither formed nor contributed to renal structures, indicating non-nephrogenic potential (Figures 5D and 5G).

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.

View Article: PubMed Central - PubMed

Affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri," Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: evangelia.papadimou@marionegri.it.

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Related in: MedlinePlus