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Journal of equine science2015; 26(3); 73-80; doi: 10.1294/jes.26.73

Population doubling level-dependent change of secreted glycosaminoglycan in equine bone marrow-derived mesenchymal stem cells.

Abstract: In regenerative medicine using transplantation of mesenchymal stem cells (MSCs), the importance of regulating the quality of MSCs has been well recognized; however, there is little information concerning the relationship between the population doubling level (PDL) and the stemness of MSCs in equine medicine. In this study, we showed that the amount of glycosaminoglycan (GAG) secreted by bone marrow-derived MSCs (BMSCs) decreases with increase of PDL. Enzymatic digestion and two-dimensional electrophoresis revealed that a main component of GAG produced by BMSCs was hyaluronan with a small amount of chondroitin sulfate. Increase of PDL downregulated the expression of MSC CD markers, including CD44, CD73, CD90, CD105, and CD146, along with loss of differentiation capacity. Thus, the effect of hyaluronan supplement to the growth medium on both expression of CD markers and the tri-lineage potential of BMSCs was evaluated. Expression of CD73 and CD90 was preserved by continuous addition of hyaluronan to the growth medium, whereas mRNA levels corresponding to CD44, CD105 and CD146 were not preserved by supplementation of hyaluronan. BMSCs subcultured with hyaluronan-supplemented growth medium to PDL-12 showed osteogenic capacity, however adipogenic and chondrogenic activities at PDL-12 were not preserved by exogenous hyaluronan. These results suggest that downregulation of CD44, CD105 and CD146 might not affect the osteogenic capacity. Taken together, the results suggested that supplementation of hyaluronan to the growth medium might be effective at maintaining the osteogenic capacity of equine BMSCs.
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Publication Date: 2015-09-30 PubMed ID: 26435680PubMed Central: PMC4591413DOI: 10.1294/jes.26.73Google Scholar: Lookup
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Summary

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The study explores the relationship between population doubling level (PDL) and the stemness of mesenchymal stem cells (MSCs) in equine medicine, highlighting how the amount of glycosaminoglycan (GAG) secreted by bone marrow-derived MSCs decreases as PDL increases. This study suggests that including hyaluronan in the growth medium may help maintain the osteogenic capacity of equine bone marrow-derived mesenchymal stem cells.

Overview of the Research

  • This research investigates the significance of maintaining the quality of MSCs when transplanting them in regenerative medicine. The focus is on determining the relationship between the population doubling level (PDL) and the stemness of MSCs in equine medicine.
  • One significant finding is that as the PDL increases, the amount of glycosaminoglycan (GAG) that is secreted by the bone marrow-derived Mesenchymal Stem Cells (BMSCs) decreases.

Glycosaminoglycan (GAG) Production

  • The study revealed that the main component of GAG produced by BMSCs was hyaluronan and supplemented with a small amount of chondroitin sulfate.
  • The research also showed that the increase in PDL leads to the decreased expression of MSC’s CD markers, among them CD44, CD73, CD90, CD105, and CD146, resulting in a loss of differentiation capacity. Differentiation capacity refers to the ability of stem cells to develop into other cell types.

Hyaluronan Supplement

  • To counter the loss of stemness, the study evaluated the effect of incorporating hyaluronan supplement to the growth medium on the expression of CD markers and the tri-lineage potential of the BMSCs.
  • It was found that continuous hyaluronan addition preserves the expression of CD73 and CD90, but the mRNA levels corresponding to CD44, CD105, and CD146 weren’t maintained.

Effects and Future Implications

  • The osteogenic capacity of BMSCs subcultured with a hyaluronan-supplemented growth medium was maintained to PDL-12, but the adipogenic and chondrogenic activities at PDL-12 were not preserved by exogenous hyaluronan.
  • The downregulation of CD44, CD105, and CD146 might not affect the osteogenic capacity, indicating the potential use of hyaluronan supplement to maintain osteogenesis of equine BMSCs.
  • In conclusion, the study suggests that maintaining the osteogenic capacity of equine BMSCs, despite an increase in PDL, might be achieved by supplementing the growth medium with hyaluronan.

Cite This Article

APA
Sasao T, Fukuda Y, Yoshida S, Miyabara S, Kasashima Y, Kuwano A, Arai K. (2015). Population doubling level-dependent change of secreted glycosaminoglycan in equine bone marrow-derived mesenchymal stem cells. J Equine Sci, 26(3), 73-80. https://doi.org/10.1294/jes.26.73

Publication

Journal of equine science
ISSN: 1340-3516
NlmUniqueID: 9503751
Country: Japan
Language: English
Volume: 26
Issue: 3
Pages: 73-80

Researcher Affiliations

Sasao, Takafumi
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Fukuda, Yuki
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Yoshida, Sayako
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Miyabara, Shihori
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
Kasashima, Yoshinori
  • Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi 320-8056, Japan.
Kuwano, Atsutoshi
  • Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi 320-8056, Japan.
Arai, Katsuhiko
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.

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Citations

This article has been cited 5 times.
  1. Harman RM, Marx C, Van de Walle GR. Translational Animal Models Provide Insight Into Mesenchymal Stromal Cell (MSC) Secretome Therapy. Front Cell Dev Biol 2021;9:654885.
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  4. Deng X, Lin B, Xiao W, Wang F, Xu P, Wang N. Specnuezhenide Alleviates Senile Osteoporosis by Activating TGR5/FXR Signaling in Bone Marrow Mesenchymal Stem Cells and RANKL-Induced Osteoclasts. Drug Des Devel Ther 2025;19:1595-1608.
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  5. Kushida C, Tamura N, Kasashima Y, Sato K, Arai K. Characterization of senescent mesenchymal stem/stromal cells derived from equine bone marrow and the effects of NANOG on the senescent phenotypes. J Vet Med Sci 2024 Sep 1;86(9):930-937.
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