top of page

Generation of PDGFRα(+) Cardioblasts from Pluripotent Stem Cells, Sci Rep. 2017 [참여저자]

최종 수정일: 9월 30일

Abstract

Isolating actively proliferating cardioblasts is the first crucial step for cardiac regeneration through cell implantation. However, the origin and identity of putative cardioblasts are still unclear. Here, we uncover a novel class of cardiac lineage cells, PDGFRα+Flk1- cardioblasts (PCBs), from mouse and human pluripotent stem cells induced using CsAYTE, a combination of the small molecules Cyclosporin A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197. This novel population of actively proliferating cells is cardiac lineage-committed but in a morphologically and functionally immature state compared to mature cardiomyocytes. Most important, most of CsAYTE-induced PCBs spontaneously differentiated into functional αMHC+ cardiomyocytes (M+CMs) and could be a potential cellular resource for cardiac regeneration.

(A) Protocol to generate PCBs from Flk1+ MPC by CsAYTE stimulation and subsequent analyses. (B) Phase-contrast images showing differentiating Flk1+ MPCs at day 6.0 incubated with control vehicle (Control), CsA, and CsAYTE. Scale bars, 100 μm. (CK) Representative FACS analyses, quantifications, and images of PDGFRα+Flk1− PCBs, PDGFRα+ Nkx2.5+ cells, and PDGFRα+ cTnT+ cells differentiated from Flk1+ MPCs at day 6.0 incubated with Control, CsA, and CsAYTE (Scale bars, 100 μm). Each group, n = 3–6. p < 0.05 and *p < 0.01 versus Con; #p < 0.05 and ##p < 0.01 versus CsA. (L) Protocol for analyses of PCB-derived cardiomyocyte differentiation in a feeder-free culture. (M and N) Representative FACS analyses and percentages of PCBs-derived cTnT+ and αMHC-GFP+ cardiomyocytes grown in feeder-free culture. Each group, n = 4–5.

조회수 0회댓글 0개

Comentários


bottom of page