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Stem cell research & therapy2018; 9(1); 75; doi: 10.1186/s13287-018-0823-3

Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells.

Abstract: Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed the release of TNF-α when exposed to LPS-stimulated monocytes similar to those cultured in FBS. ePL has the potential to be used for the expansion of MSCs before clinical application, avoiding the concerns associated with the use of FBS.
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Publication Date: 2018-03-22 PubMed ID: 29566772PubMed Central: PMC5863827DOI: 10.1186/s13287-018-0823-3Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates whether platelet lysate, specifically from horses, could be an effective alternative to fetal bovine serum (FBS) for cultivating mesenchymal stem cells (MSCs). The findings suggest that equine platelet lysate (ePL) can indeed support the growth and retain the important properties of MSCs, thereby potentially circumventing issues related to the current use of FBS.

Research Methodology

  • The research employed mesenchymal stem cells (MSCs) derived from horse bone marrow, which were cultured in two separate media conditions – one supplemented with FBS and the other with ePL.
  • The ePL was procured by a process known as apheresis, where platelets are separated from the blood. This was followed by freeze-thaw and centrifugation processes to create the lysate.
  • Proliferation rates (population doublings and doubling time) of MSCs were compared between the two media groups.
  • The overall viability of the cells, their immunophenotypic properties, and trilineage differentiation abilities were assessed.
  • To evaluate the immunomodulatory capacities of the MSCs under both conditions, they were co-cultured with monocytes stimulated by E. coli lipopolysaccharide. The level of tumor necrosis factor (TNF)-α, indicative of an inflammatory response, was measured.

Findings

  • The analysis showed that MSCs cultured with both FBS and ePL demonstrated comparable proliferation rates.
  • MSCs cultivated in ePL showed significant improvements in viability following washing compared to those cultured in FBS.
  • Expression levels of MSC markers were similar in both groups, indicating that the use of ePL did not change the cell phenotype.
  • The ability of MSCs to differentiate into three types of cells (osteocytes, adipocytes, and chondrocytes) was retained when cultured with ePL.
  • Additionally, the capacity to suppress TNF-α release when exposed to inflammatory conditions was also similar for MSCs cultured in both media types.

Implications of the Research

  • The use of FBS in stem cell culture has been linked to issues relating to xenogeneic contamination and potential alterations in cell phenotype. These issues could lead to the host immune system rejecting the transplanted cells.
  • The findings suggest that ePL might work as a viable alternative to FBS for expanding MSCs without altering their phenotype or immunomodulatory capabilities, thereby potentially eliminating the risks associated with xenogeneic contamination.

Cite This Article

APA
Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF. (2018). Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther, 9(1), 75. https://doi.org/10.1186/s13287-018-0823-3

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 9
Issue: 1
Pages: 75
PII: 75

Researcher Affiliations

Naskou, Maria C
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA.
Sumner, Scarlett M
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA.
Chocallo, Anna
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA.
Kemelmakher, Hannah
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA.
Thoresen, Merrilee
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA.
Copland, Ian
  • Emory Personalized Immunotherapy Center [EPIC], Emory University School of Medicine, 100 Woodruff Circle, Atlanta, GA, 30322, USA.
Galipeau, Jacques
  • Department of Medicine and Carbone Comprehensive Cancer Center, University of Wisconsin, 600 Highland Ave., Madison, WI, 53792, USA.
Peroni, John F
  • Department of Large Animal Medicine, Veterinary Medical Center, College of Veterinary Medicine, University of Georgia, 2200 College Station Road, Athens, GA, 30602, USA. jperoni@uga.edu.

MeSH Terms

  • Animals
  • Blood Platelets / metabolism
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / drug effects
  • Cells, Cultured
  • Culture Media, Serum-Free / chemistry
  • Culture Media, Serum-Free / pharmacology
  • Horses
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Primary Cell Culture / methods

Conflict of Interest Statement

ETHICS APPROVAL: The study protocol (IACUC approval #A2015 02–023-Y1-A1) was approved by the University of Georgia Institutional Animal Care and Committee. CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: 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.

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