title: Directed Differentiation of Human-Induced Pluripotent Stem Cells to Mesenchymal Stem Cells.
abstract: Multipotent stromal cells, also known as mesenchymal stem cells (MSCs), possess great potential to generate a wide range of cell types including endothelial cells, smooth muscle cells, bone, cartilage, and lipid cells. This protocol describes in detail how to perform highly efficient, lineage-specific differentiation of human-induced pluripotent stem cells (iPSCs) with an MSCs fate. The approach uses a clinically compliant protocol with chemically defined media, feeder-free conditions, and a CD105 positive and CD24 negative selection to achieve a single cell-based MSCs derivation from differentiating human pluripotent cells in approximately 20 days. Cells generated with this protocol express typical MSCs surface markers and undergo adipogenesis, osteogenesis, and chondrogenesis similar to adult bone marrow-derived MSCs (BM-MSCs). Nonetheless, compared with adult BM-MSCs, iPSC-MSCs display a higher proliferative capacity, up to 120 passages, without obvious loss of self-renewal potential and constitutively express MSCs surface antigens. MSCs generated with this protocol have numerous applications, including expansion to large scale cell numbers for tissue engineering and the development of cellular therapeutics. This approach has been used to rescue limb ischemia, allergic disorders, and cigarette smoke-induced lung damage and to model mesenchymal and vascular disorders of Hutchinson-Gilford progeria syndrome (HGPS).
authors.affiliation: Cardiology Division, Department of Medicine, University of Hong Kong, Hong Kong, China. qzlian@hku.hk.|Research Centre of Heart, Brain, Hormone, and Healthy Aging, Li Ka Shing Faculty of Medicine, University of Hong Kong, F/503, 5 Sassoon Road, Pokfulam, Hong Kong, China. qzlian@hku.hk.|Department of Ophthalmology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China. qzlian@hku.hk.
authors.forename: Qizhou
authors.initials: Q
authors.lastname: Lian
authors.affiliation: Cardiology Division, Department of Medicine, University of Hong Kong, Hong Kong, China.|Department of Ophthalmology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China.
authors.forename: Yuelin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Cardiology Division, Department of Medicine, University of Hong Kong, Hong Kong, China.
authors.forename: Xiaoting
authors.initials: X
authors.lastname: Liang
authors.affiliation: Department of Ophthalmology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Gao
authors.affiliation: Cardiology Division, Department of Medicine, University of Hong Kong, Hong Kong, China. hftse@hku.hk.|Research Centre of Heart, Brain, Hormone, and Healthy Aging, Li Ka Shing Faculty of Medicine, University of Hong Kong, F/503, 5 Sassoon Road, Pokfulam, Hong Kong, China. hftse@hku.hk.
authors.forename: Hung-Fat
authors.initials: HF
authors.lastname: Tse
mesh_terms: D050156:Adipogenesis; D051927:CD24 Antigen / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D000066673:Cell Self Renewal; D002478:Cells, Cultured; D020219:Chondrogenesis; D000071063:Endoglin / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000276:immunology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology; D010012:Osteogenesis
chemical_list: D051927:CD24 Antigen; C497889:CD24 protein, human; C064206:ENG protein, human; D000071063:Endoglin
keywords: Differentiation; Induced pluripotent stem cells; Mesenchymal stem cells