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Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells.

Abstract

Dynamic culture protocols have recently emerged as part of (bone) tissue engineering strategies due to their ability to represent a more physiological cell environment in vitro. Here, we described how a perfusion flow induced by a simple bioreactor system improves proliferation and osteogenic commitment of human bone marrow stromal cells. L88/5 cells were cultured in poly(methyl methacrylate) custom-milled communicating well plates, in the presence of an osteogenic cocktail containing 1α,25-dihydroxyvitamin D3, L-ascorbic acid 2-phosphate, and β-glycerophosphate. The dynamic cell culture was maintained under perfusion flow stimulation at 1 mL/min for up to 4 days and compared with a static control condition. A cell viability assay showed that the proliferation associated with the dynamic cell culture was 20% higher vs. the static condition. A significantly higher upregulation of the osteogenic markers runt-related transcription factor 2 (RUNX2), collagen type I (COL1A1), osteocalcin (BGLAP), alkaline phosphatase (ALPL), and osteopontin (SPP1) was detected when the perfusion flow stimulation was administered to the cells treated with the osteogenic cocktail. An in silico analysis showed that in the dynamic cell culture condition (i) the shear stress in the proximity of the cell layer approximates 10-3 Pa, (ii) the nutrient and the waste product concentration is more homogeneously distributed than in the static counterpart, and (iii) perfusion flow was associated with higher nutrient consumption. In summary, increased cell proliferation and enhanced early phenotype commitment indicate that dynamic cell culture conditions, delivered via bioreactor systems, produce an enhanced in vitro environment for both basic and translational research in tissue engineering and regenerative medicine.

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    ,

    Laboratory of Cellular and Molecular Engineering "S. Cavalcanti," Department of Electrical, Electronic, and Information Engineering "G. Marconi" (DEI), Alma Mater Studiorum-University of Bologna, Cesena, Italy.

    ,

    Laboratory of Cellular and Molecular Engineering "S. Cavalcanti," Department of Electrical, Electronic, and Information Engineering "G. Marconi" (DEI), Alma Mater Studiorum-University of Bologna, Cesena, Italy. Advanced Research Center on Electronic Systems (ARCES), Alma Mater Studiorum-University of Bologna, Bologna, Italy.

    ,

    Laboratory of Cellular and Molecular Engineering "S. Cavalcanti," Department of Electrical, Electronic, and Information Engineering "G. Marconi" (DEI), Alma Mater Studiorum-University of Bologna, Cesena, Italy.

    Laboratory of Cellular and Molecular Engineering "S. Cavalcanti," Department of Electrical, Electronic, and Information Engineering "G. Marconi" (DEI), Alma Mater Studiorum-University of Bologna, Cesena, Italy. Advanced Research Center on Electronic Systems (ARCES), Alma Mater Studiorum-University of Bologna, Bologna, Italy. BioEngLab, Health Science and Technology, Interdepartmental Center for Industrial Research (HST-CIRI), Alma Mater Studiorum-University of Bologna, Ozzano Emilia, Italy.

    Source

    Stem cells international 2019: 2019 pg 1304194

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31191662

    Citation

    Pasini, Alice, et al. "Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells." Stem Cells International, vol. 2019, 2019, p. 1304194.
    Pasini A, Lovecchio J, Ferretti G, et al. Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells. Stem Cells Int. 2019;2019:1304194.
    Pasini, A., Lovecchio, J., Ferretti, G., & Giordano, E. (2019). Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells. Stem Cells International, 2019, p. 1304194. doi:10.1155/2019/1304194.
    Pasini A, et al. Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells. Stem Cells Int. 2019;2019:1304194. PubMed PMID: 31191662.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells. AU - Pasini,Alice, AU - Lovecchio,Joseph, AU - Ferretti,Giulia, AU - Giordano,Emanuele, Y1 - 2019/05/02/ PY - 2018/10/31/received PY - 2019/02/04/revised PY - 2019/03/01/accepted PY - 2019/6/14/entrez PY - 2019/6/14/pubmed PY - 2019/6/14/medline SP - 1304194 EP - 1304194 JF - Stem cells international JO - Stem Cells Int VL - 2019 N2 - Dynamic culture protocols have recently emerged as part of (bone) tissue engineering strategies due to their ability to represent a more physiological cell environment in vitro. Here, we described how a perfusion flow induced by a simple bioreactor system improves proliferation and osteogenic commitment of human bone marrow stromal cells. L88/5 cells were cultured in poly(methyl methacrylate) custom-milled communicating well plates, in the presence of an osteogenic cocktail containing 1α,25-dihydroxyvitamin D3, L-ascorbic acid 2-phosphate, and β-glycerophosphate. The dynamic cell culture was maintained under perfusion flow stimulation at 1 mL/min for up to 4 days and compared with a static control condition. A cell viability assay showed that the proliferation associated with the dynamic cell culture was 20% higher vs. the static condition. A significantly higher upregulation of the osteogenic markers runt-related transcription factor 2 (RUNX2), collagen type I (COL1A1), osteocalcin (BGLAP), alkaline phosphatase (ALPL), and osteopontin (SPP1) was detected when the perfusion flow stimulation was administered to the cells treated with the osteogenic cocktail. An in silico analysis showed that in the dynamic cell culture condition (i) the shear stress in the proximity of the cell layer approximates 10-3 Pa, (ii) the nutrient and the waste product concentration is more homogeneously distributed than in the static counterpart, and (iii) perfusion flow was associated with higher nutrient consumption. In summary, increased cell proliferation and enhanced early phenotype commitment indicate that dynamic cell culture conditions, delivered via bioreactor systems, produce an enhanced in vitro environment for both basic and translational research in tissue engineering and regenerative medicine. SN - 1687-966X UR - https://www.unboundmedicine.com/medline/citation/31191662/Medium_Perfusion_Flow_Improves_Osteogenic_Commitment_of_Human_Stromal_Cells L2 - https://dx.doi.org/10.1155/2019/1304194 DB - PRIME DP - Unbound Medicine ER -