title: Expansion in microcarrier-spinner cultures improves the chondrogenic potential of human early mesenchymal stromal cells.
abstract: Cartilage tissue engineering with human mesenchymal stromal cells (hMSC) is promising for allogeneic cell therapy. To achieve large-scale hMSC propagation, scalable microcarrier-based cultures are preferred over conventional static cultures on tissue culture plastic. Yet it remains unclear how microcarrier cultures affect hMSC chondrogenic potential, and how this potential is distinguished from that of tissue culture plastic. Hence, our study aims to compare the chondrogenic potential of human early MSC (heMSC) between microcarrier-spinner and tissue culture plastic cultures.
  
  
  
  heMSC expanded on either collagen-coated Cytodex 3 microcarriers in spinner cultures or tissue culture plastic were harvested for chondrogenic pellet differentiation with empirically determined chondrogenic inducer bone morphogenetic protein 2 (BMP2). Pellet diameter, DNA content, glycosaminoglycan (GAG) and collagen II production, histological staining and gene expression of chondrogenic markers including SOX9, S100β, MMP13 and ALPL, were investigated and compared in both conditions.
  
  
  
  BMP2 was the most effective chondrogenic inducer for heMSC. Chondrogenic pellets generated from microcarrier cultures developed larger pellet diameters, and produced more DNA, GAG and collagen II per pellet with greater GAG/DNA and collagen II/DNA ratios compared with that of tissue culture plastic. Moreover, they induced higher expression of chondrogenic genes (e.g., S100β) but not of hypertrophic genes (e.g., MMP13 and ALPL). A similar trend showing enhanced chondrogenic potential was achieved with another microcarrier type, suggesting that the mechanism is due to the agitated nature of microcarrier cultures.
  
  
  
  This is the first study demonstrating that scalable microcarrier-spinner cultures enhance the chondrogenic potential of heMSC, supporting their use for large-scale cell expansion in cartilage cell therapy.
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore. Electronic address: Melissa_Lin@bti.a-star.edu.sg.
authors.forename: Youshan Melissa
authors.initials: YM
authors.lastname: Lin
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore.
authors.forename: Jessica Fang Yan
authors.initials: JF
authors.lastname: Lim
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore.
authors.forename: Jialing
authors.initials: J
authors.lastname: Lee
authors.affiliation: Experimental Fetal Medicine Group, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University Health System, Singapore.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Choolani
authors.affiliation: Experimental Fetal Medicine Group, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University Health System, Singapore; Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore; Cancer and Stem Cell Biology Program, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Jerry Kok Yen
authors.initials: JK
authors.lastname: Chan
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore.
authors.forename: Shaul
authors.initials: S
authors.lastname: Reuveny
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore. Electronic address: Steve_Oh@bti.a-star.edu.sg.
authors.forename: Steve Kah Weng
authors.initials: SK
authors.lastname: Oh
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D002356:Cartilage / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D020219:Chondrogenesis / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D004247:DNA / Q000032:analysis / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D064568:S100 Calcium Binding Protein beta Subunit / Q000096:biosynthesis; D055758:SOX9 Transcription Factor / Q000096:biosynthesis; D023822:Tissue Engineering / Q000379:methods*; D014184:Transplantation, Homologous
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D006025:Glycosaminoglycans; D064568:S100 Calcium Binding Protein beta Subunit; C579160:S100B protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D003094:Collagen; D004247:DNA; C548267:ALPL protein, human; D000469:Alkaline Phosphatase; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13
keywords: cartilage; cell therapy; chondrogenic differentiation; mesenchymal stromal cells; microcarrier