Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production

Alpa Trivedi, Byron Miyazawa, Stuart Gibb, Kristen Valanoski, Lindsay Vivona, Maximillian Lin, Daniel Potter, Mars Stone, Philip J. Norris, James Murphy, Sawyer Smith, Martin Schreiber, Shibani Pati

    Research output: Contribution to journalArticle

    Abstract

    Background: Cell based therapies, such as bone marrow derived mesenchymal stem cells (BM-MSCs; also known as mesenchymal stromal cells), are currently under investigation for a number of disease applications. The current challenge facing the field is maintaining the consistency and quality of cells especially for cell dose production for pre-clinical testing and clinical trials. Here we determine how BM-donor variability and thus the derived MSCs factor into selection of the optimal primary cell lineage for cell production and testing in a pre-clinical swine model of trauma induced acute respiratory distress syndrome. Methods: We harvested bone marrow and generated three different primary BM-MSCs from Yorkshire swine. Cells from these three donors were characterized based on (a) phenotype (morphology, differentiation capacity and flow cytometry), (b) in vitro growth kinetics and metabolic activity, and (c) functional analysis based on inhibition of lung endothelial cell permeability. Results: Cells from each swine donor exhibited varied morphology, growth rate, and doubling times. All expressed the same magnitude of standard MSC cell surface markers by flow cytometry and had similar differentiation potential. Metabolic activity and growth potential at each of the passages varied between the three primary cell cultures. More importantly, the functional potency of the MSCs on inhibition of endothelial permeability was also cell donor dependent. Conclusion: This study suggests that for production of MSCs for cell-based therapy, it is imperative to examine donor variability and characterize derived MSCs for marker expression, growth and differentiation characteristics and testing potency in application dependent assays prior to selection of the optimal cell lineage for large scale expansion and dose production.

    Original languageEnglish (US)
    Article number128
    JournalJournal of Translational Medicine
    Volume17
    Issue number1
    DOIs
    StatePublished - Apr 17 2019

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    Donor Selection
    Bone
    Bone Marrow
    Flow cytometry
    Testing
    Swine
    Functional analysis
    Cell Lineage
    Growth
    Cell- and Tissue-Based Therapy
    Mesenchymal Stromal Cells
    Growth kinetics
    Endothelial cells
    Stem cells
    Cell culture
    Permeability
    Flow Cytometry
    Assays
    Primary Cell Culture
    Adult Respiratory Distress Syndrome

    Keywords

    • Bioprocessing
    • Differentiation
    • Donor variability
    • Stem cell therapy
    • Viability

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

    Cite this

    Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production. / Trivedi, Alpa; Miyazawa, Byron; Gibb, Stuart; Valanoski, Kristen; Vivona, Lindsay; Lin, Maximillian; Potter, Daniel; Stone, Mars; Norris, Philip J.; Murphy, James; Smith, Sawyer; Schreiber, Martin; Pati, Shibani.

    In: Journal of Translational Medicine, Vol. 17, No. 1, 128, 17.04.2019.

    Research output: Contribution to journalArticle

    Trivedi, A, Miyazawa, B, Gibb, S, Valanoski, K, Vivona, L, Lin, M, Potter, D, Stone, M, Norris, PJ, Murphy, J, Smith, S, Schreiber, M & Pati, S 2019, 'Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production', Journal of Translational Medicine, vol. 17, no. 1, 128. https://doi.org/10.1186/s12967-019-1877-4
    Trivedi, Alpa ; Miyazawa, Byron ; Gibb, Stuart ; Valanoski, Kristen ; Vivona, Lindsay ; Lin, Maximillian ; Potter, Daniel ; Stone, Mars ; Norris, Philip J. ; Murphy, James ; Smith, Sawyer ; Schreiber, Martin ; Pati, Shibani. / Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production. In: Journal of Translational Medicine. 2019 ; Vol. 17, No. 1.
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    AU - Trivedi, Alpa

    AU - Miyazawa, Byron

    AU - Gibb, Stuart

    AU - Valanoski, Kristen

    AU - Vivona, Lindsay

    AU - Lin, Maximillian

    AU - Potter, Daniel

    AU - Stone, Mars

    AU - Norris, Philip J.

    AU - Murphy, James

    AU - Smith, Sawyer

    AU - Schreiber, Martin

    AU - Pati, Shibani

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    AB - Background: Cell based therapies, such as bone marrow derived mesenchymal stem cells (BM-MSCs; also known as mesenchymal stromal cells), are currently under investigation for a number of disease applications. The current challenge facing the field is maintaining the consistency and quality of cells especially for cell dose production for pre-clinical testing and clinical trials. Here we determine how BM-donor variability and thus the derived MSCs factor into selection of the optimal primary cell lineage for cell production and testing in a pre-clinical swine model of trauma induced acute respiratory distress syndrome. Methods: We harvested bone marrow and generated three different primary BM-MSCs from Yorkshire swine. Cells from these three donors were characterized based on (a) phenotype (morphology, differentiation capacity and flow cytometry), (b) in vitro growth kinetics and metabolic activity, and (c) functional analysis based on inhibition of lung endothelial cell permeability. Results: Cells from each swine donor exhibited varied morphology, growth rate, and doubling times. All expressed the same magnitude of standard MSC cell surface markers by flow cytometry and had similar differentiation potential. Metabolic activity and growth potential at each of the passages varied between the three primary cell cultures. More importantly, the functional potency of the MSCs on inhibition of endothelial permeability was also cell donor dependent. Conclusion: This study suggests that for production of MSCs for cell-based therapy, it is imperative to examine donor variability and characterize derived MSCs for marker expression, growth and differentiation characteristics and testing potency in application dependent assays prior to selection of the optimal cell lineage for large scale expansion and dose production.

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