Expansion in microcarrier-spinner cultures improves the chondrogenic potential of human early mesenchymal stromal cells

Autor: Jerry Kok Yen Chan, Jessica Fang Yan Lim, Youshan Melissa Lin, Shaul Reuveny, Steve Oh, Mahesh Choolani, Jialing Lee
Rok vydání: 2015
Předmět:
0301 basic medicine
Cancer Research
Immunology
Cell Culture Techniques
Cell- and Tissue-Based Therapy
Bone Morphogenetic Protein 2
S100 Calcium Binding Protein beta Subunit
Mesenchymal Stem Cell Transplantation
Bone morphogenetic protein 2
03 medical and health sciences
Tissue culture
0302 clinical medicine
Chondrocytes
Tissue engineering
Matrix Metalloproteinase 13
medicine
Immunology and Allergy
Humans
Transplantation
Homologous
Genetics (clinical)
Cells
Cultured
Cell Proliferation
Glycosaminoglycans
Transplantation
Tissue Engineering
Chemistry
Cartilage
Mesenchymal stem cell
Microcarrier
Cell Differentiation
Mesenchymal Stem Cells
SOX9 Transcription Factor
Cell Biology
DNA
Chondrogenesis
Alkaline Phosphatase
Cell biology
030104 developmental biology
medicine.anatomical_structure
Oncology
Cell culture
030220 oncology & carcinogenesis
Collagen
Zdroj: Cytotherapy. 18(6)
ISSN: 1477-2566
Popis: Background aims 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. Methods 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. Results 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. Conclusions 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.
Databáze: OpenAIRE
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