title: Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs).
abstract: Stem cells (SCs) are a promising approach to regenerative medicine, with the potential to treat numerous orthopedic disorders, including osteo-degenerative diseases. The development of human-induced pluripotent stem cells (hiPSCs) has increased the potential of SCs for new treatments. However, current methods of differentiating hiPSCs into chondrocyte-like cells are suboptimal and better methods are needed. The aim of the present study was to assess four different chondrogenic differentiation protocols to identify the most efficient method of generating hiPSC-derived chondrocytes. For this study, hiPSCs were obtained from primary hu﻿man dermal fibroblasts (PHDFs) and differentiated into chondrocyte-like cells using four different protocols: 1) monolayer culture with defined growth factors (GF); 2) embryoid bodies (EBs) in a chondrogenic medium with TGF-β3 cells; 3) EBs in chondrogenic medium conditioned with human chondrocytes (HC-402-05a cell line) and 4) EBs in chondrogenic medium conditioned with human chondrocytes and supplemented with TGF-β3. The cells obtained through these four protocols were evaluated and compared at the mRNA and protein levels. Although chondrogenic differentiation of hiPSCs was successfully achieved with all of these protocols, the two fastest and most cost-effective methods were the monolayer culture with GFs and the medium conditioned with human chondrocytes. Both of these methods are superior to other available techniques. The main advantage of the conditioned medium is that the technique is relatively simple and inexpensive while the directed method (i.e., monolayer culture with GFs) is faster than any protocol described to date because it is does not require additional steps such as EB formation.
authors.affiliation: Radiobiology Lab, Greater Poland Cancer Centre, 61- 866, Poznan, Poland.|Department of Electroradiology, Poznan University of Medical Sciences, 61-866, Poznan, Poland.
authors.forename: Wiktoria Maria
authors.initials: WM
authors.lastname: Suchorska
authors.affiliation: Radiobiology Lab, Greater Poland Cancer Centre, 61- 866, Poznan, Poland. ewelina.augustyniak@wco.pl.|The Postgraduate School of Molecular Medicine, Medical University of Warsaw, 02-091, Warsaw, Poland. ewelina.augustyniak@wco.pl.
authors.forename: Ewelina
authors.initials: E
authors.lastname: Augustyniak
authors.affiliation: Department of Orthopedics and Traumatology, Poznan University of Medical Sciences, 61-545, Poznan, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Richter
authors.affiliation: Department of Orthopedics and Traumatology, Poznan University of Medical Sciences, 61-545, Poznan, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Trzeciak
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects / Q000235:genetics; D017077:Culture Media, Conditioned / Q000494:pharmacology; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008856:Microscopy, Fluorescence; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D055755:SOXD Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; C486270:SOX6 protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D055755:SOXD Transcription Factors
keywords: Cell differentiation; Chondrocytes; Human induced pluripotent stem cells; Regenerative medicine