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

• Type of Material: Article
• Authors: Courtney M. Caroti, Emory University; Hyunhee Ahn, Emory University; Hector F. Salazar, Emory University; Giji Joseph, Emory University; Sitara B. Sankar, Georgia Institute of Technology; Nick Willett, Emory University; Levi B. Wood, Georgia Institute of Technology; W Robert Taylor, Emory University; Alicia N. Lyle, Emory University
• Language: English
• Date: 2017-10-17
• Publisher: Nature Publishing Group: Open Access Journals - Option C
• Publication Version: Final Published Version
• Copyright Statement: © 2017 The Author(s).
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41598-017-13477-y
• Title of Journal or Parent Work: Scientific Reports
• ISSN: 2045-2322
• Volume: 7
• Issue: 1
• Start Page: 13334
• End Page: 13334
• Grant/Funding Information: 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: INBRED MICE; Science & Technology - Other Topics; THERAPY; DIFFERENTIATION; Science & Technology; MOUSE BONE-MARROW; EXPRESSION; MYOCARDIAL-INFARCTION; PROLIFERATION; STROMAL CELLS; LIMB ISCHEMIA; GROWTH; Multidisciplinary Sciences
• Research Categories: Engineering, Mechanical; Health Sciences, General; Engineering, Biomedical

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.