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Author Notes:

Correspondence and requests for materials should be addressed to A.N.L. (email: alicia.lyle@emory.edu)

C.M.C. and H.A. contributed equally to this study.

C.M.C. conducted cell culture studies, flow cytometry, senescence staining, and proliferation and differentiation studies, as well as data analysis and manuscript preparation.

H.A. contributed to flow cytometry, CFU assay, senescence analysis, data analysis and manuscript preparation.

H.F.S. conducted cell culture studies, differentiation and proliferation studies, and performed data analysis.

G.J. conducted histology and cell staining for encapsulated MSCs.

S.S. assisted with the multiplex cytokine analysis.

L.B.W. conducted the multiplexed analysis of cytokines and phospho-proteins.

A.N.L., W.R.T., and N.J.W. supervised this research.

A.N.L. designed and contributed to all aspects of study.

All authors reviewed the manuscript.

We thank Dr. Jacques Galipeau (Department of Hematology and Medical Oncology, Emory University) and Dr. Alexandra Peister (Department of Biology, Morehouse College) for their helpful discussion and insights.

We also thank the Emory University School of Medicine Core Facilities including the Microscopy in Medicine Core, supported by NIH PPG # 5P01HL095070, and the Core Facility for Flow Cytometry.

The authors declare that they have no competing interests.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Subjects:

Research Funding:

This work was supported by a K99/R00 grant awarded to Alicia N. Lyle from the National Heart, Lung and Blood Institute (K99/R00 HL119567) and startup funds from the Woodruff School of Mechanical Engineering at the Georgia Institute of Technology to Levi B. Wood.

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • MOUSE BONE-MARROW
  • STROMAL CELLS
  • MYOCARDIAL-INFARCTION
  • LIMB ISCHEMIA
  • INBRED MICE
  • GROWTH
  • DIFFERENTIATION
  • EXPRESSION
  • THERAPY
  • PROLIFERATION

A Novel Technique for Accelerated Culture of Murine Mesenchymal Stem Cells that Allows for Sustained Multipotency

Tools:

Journal Title:

Scientific Reports

Volume:

Volume 7, Number 1

Publisher:

, Pages 13334-13334

Type of Work:

Article | Final Publisher PDF

Abstract:

Bone marrow derived mesenchymal stem cells (MSCs) are regularly utilized for translational therapeutic strategies including cell therapy, tissue engineering, and regenerative medicine and are frequently used in preclinical mouse models for both mechanistic studies and screening of new cell based therapies. Current methods to culture murine MSCs (mMSCs) select for rapidly dividing colonies and require long-term expansion. These methods thus require months of culture to generate sufficient cell numbers for feasibility studies in a lab setting and the cell populations often have reduced proliferation and differentiation potential, or have become immortalized cells. Here we describe a simple and reproducible method to generate mMSCs by utilizing hypoxia and basic fibroblast growth factor supplementation. Cells produced using these conditions were generated 2.8 times faster than under traditional methods and the mMSCs showed decreased senescence and maintained their multipotency and differentiation potential until passage 11 and beyond. Our method for mMSC isolation and expansion will significantly improve the utility of this critical cell source in pre-clinical studies for the investigation of MSC mechanisms, therapies, and cell manufacturing strategies.

Copyright information:

© 2017 The Author(s).

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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