A R Tan1, E Alegre-Aguarón1, G D O'Connell2, C D VandenBerg3, R K Aaron4, G Vunjak-Novakovic1, J Chloe Bulinski5, G A Ateshian6, C T Hung7. 1. Department of Biomedical Engineering, Columbia University, 1210 Amsterdam Ave, New York, NY 10027, USA. 2. Department of Mechanical Engineering, University of California, Berkeley, 5122 Etcheverry Hall, Berkeley, CA 94720, USA. 3. Department of Orthopaedic Surgery, St. Luke's-Roosevelt Hospital Center, 1000 10th Ave, New York, NY 10019, USA. 4. Department of Orthopaedic Surgery, Brown University, 100 Butler Drive, Providence, RI 02906, USA. 5. Department of Biological Sciences, Columbia University, 1212 Amsterdam Ave, New York, NY 10027, USA. 6. Department of Biomedical Engineering, Columbia University, 1210 Amsterdam Ave, New York, NY 10027, USA; Department of Mechanical Engineering, Columbia University, 500 W. 120th St, New York, NY 10027, USA. 7. Department of Biomedical Engineering, Columbia University, 1210 Amsterdam Ave, New York, NY 10027, USA. Electronic address: cth6@columbia.edu.
Abstract
OBJECTIVE: Galvanotaxis, the migratory response of cells in response to electrical stimulation, has been implicated in development and wound healing. The use of mesenchymal stem cells (MSCs) from the synovium (synovium-derived stem cells, SDSCs) has been investigated for repair strategies. Expansion of SDSCs is necessary to achieve clinically relevant cell numbers; however, the effects of culture passage on their subsequent cartilaginous extracellular matrix production are not well understood. METHODS: Over four passages of SDSCs, we measured the expression of cell surface markers (CD31, CD34, CD49c, CD73) and assessed their migratory potential in response to applied direct current (DC) electric field. Cells from each passage were also used to form micropellets to assess the degree of cartilage-like tissue formation. RESULTS: Expression of CD31, CD34, and CD49c remained constant throughout cell expansion; CD73 showed a transient increase through the first two passages. Correspondingly, we observed that early passage SDSCs exhibit anodal migration when subjected to applied DC electric field strength of 6 V/cm. By passage 3, CD73 expression significantly decreased; these cells exhibited cell migration toward the cathode, as previously observed for terminally differentiated chondrocytes. Only late passage cells (P4) were capable of developing cartilage-like tissue in micropellet culture. CONCLUSIONS: Our results show cell priming protocols carried out for four passages selectively differentiate stem cells to behave like chondrocytes, both in their motility response to applied electric field and their production of cartilaginous tissue.
OBJECTIVE: Galvanotaxis, the migratory response of cells in response to electrical stimulation, has been implicated in development and wound healing. The use of mesenchymal stem cells (MSCs) from the synovium (synovium-derived stem cells, SDSCs) has been investigated for repair strategies. Expansion of SDSCs is necessary to achieve clinically relevant cell numbers; however, the effects of culture passage on their subsequent cartilaginous extracellular matrix production are not well understood. METHODS: Over four passages of SDSCs, we measured the expression of cell surface markers (CD31, CD34, CD49c, CD73) and assessed their migratory potential in response to applied direct current (DC) electric field. Cells from each passage were also used to form micropellets to assess the degree of cartilage-like tissue formation. RESULTS: Expression of CD31, CD34, and CD49c remained constant throughout cell expansion; CD73 showed a transient increase through the first two passages. Correspondingly, we observed that early passage SDSCs exhibit anodal migration when subjected to applied DC electric field strength of 6 V/cm. By passage 3, CD73 expression significantly decreased; these cells exhibited cell migration toward the cathode, as previously observed for terminally differentiated chondrocytes. Only late passage cells (P4) were capable of developing cartilage-like tissue in micropellet culture. CONCLUSIONS: Our results show cell priming protocols carried out for four passages selectively differentiate stem cells to behave like chondrocytes, both in their motility response to applied electric field and their production of cartilaginous tissue.
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