title: Chemically defined serum-free conditions for cartilage regeneration from human embryonic stem cells.
abstract: OBJECTIVE
  The aim of this study was to improve a method that induce cartilage differentiation of human embryoid stem cells (hESCs) in vitro, and test the effect of in vivo environments on the further maturation of hESCs derived cells.
  
  
  METHODS
  Embryoid bodies (EBs) formed from hESCs, with serum-free KSR-based medium and mesodermal specification related factors, CHIR, and Noggin for first 8days. Then cells were digested and cultured as micropellets in serum-free KSR-based chondrogenic medium that was supplemented with PDGF-BB, TGF β3, BMP4 in sequence for 24days. The morphology, FACS, histological staining as well as the expression of chondrogenic specific genes were detected in each stage, and further in vivo experiments, cell injections and tissue transplantations, further verified the formation of chondrocytes.
  
  
  RESULTS
  We were able to obtain chondrocyte/cartilage from hESCs using serum-free KSR-based conditioned medium. qPCR analysis showed that expression of the chondroprogenitor genes and the chondrocyte/cartilage matrix genes. Morphology analysis demonstrated we got PG+COL2+COL1-particles. It indicated we obtained hyaline cartilage-like particles. 32-Day differential cells were injected subcutaneous. Staining results showed grafts developed further mature in vivo. But when transplanted in subrenal capsule, their effect was not good as in subcutaneous. Microenvironment might affect the cartilage formation.
  
  
  CONCLUSIONS
  The results of this study provide an absolute serum-free and efficient approach for generation of hESC-derived chondrocytes, and cells will become further maturation in vivo. It provides evidence and technology for the hypothesis that hESCs may be a promising therapy for the treatment of cartilage disease.
authors.affiliation: Experimental Center of Pathogenobiology Immunology Cytobiology and Genetics, Basic Medical College, Jilin University, Changchun, PR China; Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, PR China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Yang
authors.affiliation: Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, PR China.
authors.forename: Shubin
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Orthopaedic Surgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, PR China.
authors.forename: Changzhao
authors.initials: C
authors.lastname: Gao
authors.affiliation: Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, PR China; Department of Regenerative Medicine, School of Pharmaceutical Science, Jilin University, Changchun, PR China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Regenerative Medicine, School of Pharmaceutical Science, Jilin University, Changchun, PR China.
authors.forename: Yulai
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, USA. Electronic address: rockman123456@sina.com.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Liu
authors.affiliation: Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, PR China. Electronic address: cai_jinglei@gibh.ac.cn.
authors.forename: Jinglei
authors.initials: J
authors.lastname: Cai
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D000818:Animals; D000077214:Becaplermin; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002356:Cartilage / Q000166:cytology / Q000502:physiology*; D064236:Cartilage Oligomeric Matrix Protein / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology*; D005434:Flow Cytometry; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D016133:Polymerase Chain Reaction; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D012038:Regeneration / Q000187:drug effects*; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology
chemical_list: D043925:Angiogenesis Inducing Agents; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D064236:Cartilage Oligomeric Matrix Protein; D016895:Culture Media, Serum-Free; D020574:Proto-Oncogene Proteins c-sis; D053782:Transforming Growth Factor beta3; D000077214:Becaplermin
keywords: Cartilage; Differentiation; Serum-free; Stem cells