Format

Send to

Choose Destination
Gene. 2020 Jan 5;722:144058. doi: 10.1016/j.gene.2019.144058. Epub 2019 Sep 5.

Hypothermia and nutrient deprivation alter viability of human adipose-derived mesenchymal stem cells.

Author information

1
Department of Physical Medicine &Rehabilitation, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America; Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America; Department of Biochemistry & Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
2
Department of Physical Medicine &Rehabilitation, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America. Electronic address: Qu.wenchun@mayo.edu.
3
Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America; Department of Biochemistry & Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
4
Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America; Center for Regenerative Medicine, Mayo Clinic, Rochester, MN, United States of America; Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, MN, United States of America.
5
Department of Physical Medicine &Rehabilitation, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
6
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States of America.
7
Department of Orthopedic Surgery, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America; Department of Biochemistry & Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America. Electronic address: vanwijnen.andre@mayo.edu.

Abstract

PURPOSE:

Adipose-derived mesenchymal stem cells (MSCs) are attractive biological agents in regenerative medicine. To optimize cell therapies, it is necessary to determine the most effective delivery method for MSCs. Therefore, we evaluated the biological properties of MSCs after exposure to various temperatures to define optimal storage conditions prior to therapeutic delivery of MSCs.

DESIGN:

Prospective observational study.

METHODS AND MATERIALS:

Adherent and non-adherent MSCs were incubated at multiple temperatures (i.e., 4, 23 and 37 °C) in Lactated Ringers (LR) solution lacking essential cell growth ingredients, or in culture media which is optimized for cell growth. Cells were assessed either after the temperature changes (4 h) or after recovery (24 h). Metabolic activity of MSCs, cell number and expression of representative mRNA biomarkers were evaluated to assess the biological effects of temperature. We monitored changes in mRNAs expression related to cytoprotective- or stress-related responses (e.g., FOS, JUN, ATF1, ATF4, EGR1, EGR2, MYC), proliferation (e.g., HIST2H4, CCNB2), and extracellular matrix production (ECM; e.g., COL3A1, COL1A1) by quantitative real time reverse-transcriptase polymerase chain reaction (RT-qPCR) analysis.

RESULTS:

Our study demonstrates that storing MSCs in Lactated Ringers (LR) solution for 4 h decreases cell number and metabolic activity. The number of viable MSCs decreased significantly when cultured at physiological temperature (37 °C) and severe hypothermia (4 °C), while cells grown at ambient temperature (23 °C) exhibited the least detrimental effects. There were no appreciable biological differences in mRNA markers for proliferation or ECM deposition at any of the temperatures. However, biomarkers related to cytoprotective- or stress-responses were selectively elevated depending on temperature or media type (i.e., LR versus standard media).

CONCLUSION:

The biological impact of nutrient-free media and temperature changes after 4 h exposure persists after a 24 h recovery period. Hence, storage temperature and media conditions should be optimized to improve effective dosing of MSCs.

KEYWORDS:

Basic science; Cell stress; Connective tissue diseases; Hypothermia; Hypoxia; Mesenchymal stem cell; Musculoskeletal conditions; Quality improvement and patient safety; Stem cell therapy

PMID:
31494240
DOI:
10.1016/j.gene.2019.144058
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center