Early induction of a prechondrogenic population allows efficient generation of stable chondrocytes from human induced pluripotent stem cells.
Regeneration of human cartilage is inherently inefficient; an abundant autologous source, such as human induced pluripotent stem cells (hiPSCs), is therefore attractive for engineering cartilage. We report a growth factor-based protocol for differentiating hiPSCs into articular-like chondrocytes (hiChondrocytes) within 2 weeks, with an overall efficiency >90%. The hiChondrocytes are stable and comparable to adult articular chondrocytes in global gene expression, extracellular matrix production, and ability to generate cartilage tissue in vitro and in immune-deficient mice. Molecular characterization identified an early SRY (sex-determining region Y) box (Sox)9(low) cluster of differentiation (CD)44(low)CD140(low) prechondrogenic population during hiPSC differentiation. In addition, 2 distinct Sox9-regulated gene networks were identified in the Sox9(low) and Sox9(high) populations providing novel molecular insights into chondrogenic fate commitment and differentiation. Our findings present a favorable method for generating hiPSC-derived articular-like chondrocytes. The hiChondrocytes are an attractive cell source for cartilage engineering because of their abundance, autologous nature, and potential to generate articular-like cartilage rather than fibrocartilage. In addition, hiChondrocytes can be excellent tools for modeling human musculoskeletal diseases in a dish and for rapid drug screening.
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
Jieun
J
Lee
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
Sarah E B
SE
Taylor
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
Piera
P
Smeriglio
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
Janice
J
Lai
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
William J
WJ
Maloney
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA.
Fan
F
Yang
*Department of Orthopaedic Surgery, Department of Mechanical Engineering, and Department of Bioengineering, Stanford University School of Medicine, Stanford, California, USA nbhutani@stanford.edu.
Nidhi
N
Bhutani
D000328:Adult; D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism / Q000502:physiology*; D020219:Chondrogenesis / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D008297:Male; D051379:Mice; D012038:Regeneration / Q000502:physiology*; D055758:SOX9 Transcription Factor / Q000378:metabolism
D055758:SOX9 Transcription Factor
cartilage regeneration; hiPSC-derived chondrocytes; prechondrogenic population