title: Pluripotent Conversion of Muscle Stem Cells Without Reprogramming Factors or Small Molecules.
abstract: Muscle derived stem cells (MDSCs) are multipotent stem cells that can differentiate into several lineages including skeletal muscle precursor cells. Here, we show that MDSCs from myostatin null mice (Mstn (-/-) ) can be readily induced into pluripotent stem cells without using reprogramming factors. Microarray studies revealed a strong upregulation of markers like Leukemia Inhibitory factor (LIF) and Leukemia Inhibitory factor receptor (LIFR) in Mstn (-/-) MDSCs as compared to wild type MDSCs (WT-MDSCs). Furthermore when cultured in mouse embryonic stem cell media with LIF for 95 days, Mstn (-/-) MDSCs formed embryonic stem cell (ES) like colonies. We termed such ES like cells as the culture-induced pluripotent stem cells (CiPSC). CiPSCs from Mstn (-/-) MDSCs were phenotypically similar to ESCs, expressed high levels of Oct4, Nanog, Sox2 and SSEA-1, maintained a normal karyotype. Furthermore, CiPSCs formed embryoid bodies and teratomas when injected into immunocompromised mice. In addition, CiPSCs differentiated into somatic cells of all three lineages. We further show that culturing in ES cell media, resulted in hypermethylation and downregulation of BMP2 in Mstn(-/-) MDSCs. Western blot further confirmed a down regulation of BMP2 signaling in Mstn (-/-) MDSCs in supportive of pluripotent reprogramming. Given that down regulation of BMP2 has been shown to induce pluripotency in cells, we propose that lack of myostatin epigenetically reprograms the MDSCs to become pluripotent stem cells. Thus, here we report the successful establishment of ES-like cells from adult stem cells of the non-germline origin under culture-induced conditions without introducing reprogramming genes.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60, Nanyang Drive, Singapore, 637551, Singapore. Bipasha.bose@gmail.com.|Stem Cell and Tissue Engineering Division, Yenepoya Research Center, Yenepoya University, University Road, Derlakatte, Mangalore, Karnataka, 575018, India. Bipasha.bose@gmail.com.
authors.forename: Bipasha
authors.initials: B
authors.lastname: Bose
authors.affiliation: School of Biological Sciences, Nanyang Technological University, 60, Nanyang Drive, Singapore, 637551, Singapore. shenoy2000@yahoo.com.|Stem Cell and Tissue Engineering Division, Yenepoya Research Center, Yenepoya University, University Road, Derlakatte, Mangalore, Karnataka, 575018, India. shenoy2000@yahoo.com.
authors.forename: Sudheer
authors.initials: S
authors.lastname: Shenoy P
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D003470:Culture Media / Q000494:pharmacology*; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007621:Karyotyping; D053605:Leukemia Inhibitory Factor / Q000235:genetics / Q000378:metabolism; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit / Q000235:genetics / Q000378:metabolism; D016256:Lewis X Antigen / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D046228:Microarray Analysis; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D055435:Myostatin / Q000172:deficiency* / Q000235:genetics; D000071317:Nanog Homeobox Protein / Q000235:genetics / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D015415:Biomarkers; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D003470:Culture Media; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D016256:Lewis X Antigen; C507838:Lif protein, mouse; C507841:Lifr protein, mouse; C525912:Mstn protein, mouse; D055435:Myostatin; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse
keywords: BMP2; Induced pluripotency; MAP kinase; Multipotency; Muscle derived stem cells; Myostatin null; Smad