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Veterinary research communications2018; 42(3); 171-181; doi: 10.1007/s11259-018-9720-6

Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity.

Abstract: Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0 × 10 cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.
Publication Date: 2018-05-02 PubMed ID: 29721754DOI: 10.1007/s11259-018-9720-6Google Scholar: Lookup
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  • Journal Article

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.

The research assesses the potential of using a three-dimensional culture system to enhance the properties of stem cells derived from horse adipose tissue, which could subsequently improve treatment for horse fractures.

Understanding Stem Cells and their Potential Application

  • The study is primarily concerned with mesenchymal stem cells (MSCs), which are a type of cell that can generate into a variety of cell types. These stem cells, in particular, are derived from equine adipose tissue (or fat tissue from horses) and are referred to as eAD-MSCs.
  • Scientists believe MSCs could be valuable in tissue engineering and cell therapy fields, which include efforts to heal or replace damaged tissues and organs.
  • The focus here is on harnessing these cells to better treat horse fractures. Scientists are specifically investigating whether a three-dimensional (3D) culture system might enhance the properties of these cells.

Methodology: 3D Spheroid Formation

  • The eAD-MSCs are successfully extracted from a horse’s tailhead. These cells are then put through a process known as the hanging drop method, which generates a spheroid formation (a spherical mass) of these cells.
  • This spheroid formation occurs over three days, following which the scientists performed Quantitative Real-Time PCR tests to analyze the levels of gene expression within the cells.

Outcomes: Changes in Gene Expression Levels

  • The results demonstrate that the spheroid formation of the cells led to heightened expression levels of pluripotent markers, PGE production, and certain angiogenesis-inducing (blood vessel forming) factors like VEGF, IL-6, IL-8, and IL-18.
  • Levels of Ki-67, a protein associated with cell growth and proliferation, also significantly increased in the spheroid formation compared to when the cells were cultured in a monolayer (a single, flat layer).
  • The spheroid formation also demonstrated superior expression of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1, and osteocyte-specific markers such as RUNX2 and osteocalcin, indicating better potential for differentiation into bone cells or osteocytes.

Conclusion

The findings suggest that the three-dimensional spheroid formation of equine adipose tissue-derived MSCs, achieved via the hanging drop method, can raise the expression of angiogenesis-inducing and bone morphogenesis-inducing factors within these cells, which might enhance their therapeutic potential.

Cite This Article

APA
Park MJ, Lee J, Byeon JS, Jeong DU, Gu NY, Cho IS, Cha SH. (2018). Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity. Vet Res Commun, 42(3), 171-181. https://doi.org/10.1007/s11259-018-9720-6

Publication

ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 42
Issue: 3
Pages: 171-181

Researcher Affiliations

Park, Mi Jeong
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Lee, Jienny
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Byeon, Jeong Su
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Jeong, Da-Un
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Gu, Na-Yeon
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Cho, In-Soo
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea.
Cha, Sang-Ho
  • Viral Disease Research Division, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si, Gyeongsangbuk-do, 39660, Republic of Korea. virusmania@korea.kr.

MeSH Terms

  • Adipose Tissue / cytology
  • Animals
  • Cell Culture Techniques / veterinary
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Female
  • Horses / physiology
  • Male
  • Mesenchymal Stem Cells / cytology
  • Spheroids, Cellular / physiology

Grant Funding

  • M-1543083-2014-17-01 / Animal and Plant Quarantine Agency

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