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Directing human pluripotent stem cells toward the skeletal muscle lineage
Author(s) -
Tran Tuan,
Peterson Cory,
Pyle April D.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.705.5
Subject(s) - embryoid body , induced pluripotent stem cell , embryonic stem cell , biology , germ layer , microbiology and biotechnology , skeletal muscle , progenitor cell , p19 cell , stem cell , directed differentiation , cellular differentiation , regenerative medicine , mesenchymal stem cell , cell type , myogenesis , myocyte , cell , anatomy , genetics , gene
Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) hold great promise as a potential source of cells for use in therapeutic medicine due to their ability to differentiate into cells from each germ layer and continuously self‐renew. Despite this potential, there remains great variation in both methodology and efficiency of generating mesodermal lineages from either hESCs or hiPSCs. Of particular difficulty has been the development of reliable protocols to efficiently differentiate skeletal muscle lineages. Although CD73 + mesenchymal cells have been obtained through sequential culturing and cell sorting methods, skeletal muscle cell yield was low and the protocol requires media for neuronal growth. Therefore we are currently establishing an alternative approach for directing differentiation of hESCs and hiPSCs toward the muscle lineage. To this end, when embryoid bodies (EB) derived from hESCs and hiPSCs were differentiated after 24 days, maximal enrichment for early stage or progenitor muscle markers was achieved compared to those differentiated up to 30 days. Subsequently, dissociated cells from EBs at each time point were sorted for either NCAM or M‐Cadherin and expanded in standard culture conditions for promoting muscle cell growth. Both cell types were confirmed to express Pax3 via RT‐PCR. Establishment of standard protocols for the reliable generation of muscle progenitor cells from PSCs will be important in future experiments for assaying the differentiation potential of Duchenne Muscular Dystrophy (DMD) patient‐specific hiPSCs as well as for identifying the muscle cell type(s) needed for efficacious cell therapy.