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April 27, 2022

Single-cell RNA sequencing of hiPSC-derived muscle progenitor cells identifies key factors of proliferation

Transcriptomic Atlas of hiPSC-derived muscle progenitor cells cultures. (A) Schematic representation of the myogenic induction protocol. Cells were analyzed after 80 d of differentiation. CHIR, CHIR99021; HS, horse serum; SB, SB431542. (B) Immunohistochemical analysis of PAX7 (green), MYOD1 (white), MYH (MYOSIN HEAVY CHAIN, green), and DAPI (blue) at day 80 of the differentiation. Scale bar, 200 μm. (C) Quantification of PAX7 and MYOD in dissociated hiPSC-MuPCs at day 80 of the differentiation. Data represent the mean of three independent experiments. (B, D) A transcriptomic atlas of all cells dissociated from the cells in (B). (E) Single-cell expression of the selected markers. (F) Violin plots showing expression clusters of the selected markers. (G) A Gene Ontology analysis (biological processes) for differentially expressed genes up-regulated in each cluster. Credit: Life Science Alliance (2022). DOI: 10.26508/lsa.202101312
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Transcriptomic Atlas of hiPSC-derived muscle progenitor cells cultures. (A) Schematic representation of the myogenic induction protocol. Cells were analyzed after 80 d of differentiation. CHIR, CHIR99021; HS, horse serum; SB, SB431542. (B) Immunohistochemical analysis of PAX7 (green), MYOD1 (white), MYH (MYOSIN HEAVY CHAIN, green), and DAPI (blue) at day 80 of the differentiation. Scale bar, 200 μm. (C) Quantification of PAX7 and MYOD in dissociated hiPSC-MuPCs at day 80 of the differentiation. Data represent the mean of three independent experiments. (B, D) A transcriptomic atlas of all cells dissociated from the cells in (B). (E) Single-cell expression of the selected markers. (F) Violin plots showing expression clusters of the selected markers. (G) A Gene Ontology analysis (biological processes) for differentially expressed genes up-regulated in each cluster. Credit: Life Science Alliance (2022). DOI: 10.26508/lsa.202101312

A group led by Dr. Minas Nalbandian has established an atlas of human pluripotent stem cell-derived muscle progenitor cells (hiPSC-MuPCs) and found the heterogeneity in hiPSC-MuPCs.

The research group has been developing transplantation therapy of hiPSC-MuPCs as a treatment for skeletal muscle diseases such as Duchenne (DMD). However, it was unclear whether hiPSC-MuPCs has a uniform cell population or not.

In this study, the group performed single-cell RNA sequencing (scRNA-seq) of hiPSC-MuPCs cultures to study the cell heterogeneity of the myogenic subset of cells and found four clusters of cells: noncycling progenitors, cycling, committed, and myocytes. Furthermore, they found FGFR4 and CD36 to be useful markers for separating these subpopulations and demonstrated that the FGFR4-positive cell population has a higher regenerative capacity.

The findings were published in Life Science Alliance on 22 April 2022.

More information: Minas Nalbandian et al, Single-cell RNAseq reveals heterogeneity in hiPSC-MuPCs and E2F as a key regulator of proliferation, Life Science Alliance (2022).

Provided by Kyoto University

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