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Home / Equine Research Bank / Stem Cells Dev / Temporal analysis of equine bone marrow aspirate during esta...
Stem cells and development2009; 19(2); 269-282; doi: 10.1089/scd.2009.0091

Temporal analysis of equine bone marrow aspirate during establishment of putative mesenchymal progenitor cell populations.

Abstract: Mesenchymal progenitor cells (MPCs) are often characterized using surface markers after expansion and treatment in culture. There are no studies directly comparing gene and protein markers in undifferentiated samples during the very early phases of culture. The goal of this study was to evaluate temporal gene and protein expression changes during establishment of equine MPC cultures. Bone marrow aspirate was obtained from 35 horses and processed by density gradient centrifugation. In freshly isolated bone marrow, mononuclear cells had variable expression of CD44, CD11a/CD18, CD90, and CD45RB cell surface molecules. After 2 h of culture, bone marrow mononuclear cells had a phenotype of CD44(hi), CD29(hi), CD90(lo), CD11a/CD18(hi), and CD45RB(lo). Isolated mononuclear cells were analyzed by flow cytometry and RT-qPCR at 2, 7, 14, 21, and 30 days of culture. At all culture time points, gene expression was in agreement with cell surface protein expression. In established cultures of MPCs, cells remained robustly positive for CD44 and CD29. The proportion of positive cells and the mean fluorescence intensity of positive cells increased in CD90 expression as MPC cultures became more homogeneous. Inversely, the population of cells in culture decreased expression of CD11a/CD18 and CD45RB molecules over time. The decreased expression of the latter molecules makes these useful negative markers of established MPC cultures under normal expansion conditions. The results of this study demonstrate numerous dynamic changes in cell surface molecule expression during early establishment of MPC populations, which may aid to improve MPC isolation methods for research or therapeutic applications.
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Publication Date: 2009-07-17 PubMed ID: 19604071PubMed Central: PMC3138180DOI: 10.1089/scd.2009.0091Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

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 paper reveals the changes in gene and protein expression during the early phase of horse mesenchymal progenitor cell (MPC) culture establishment. The study deepens our understanding of this cell population, with potential implications for improving research and therapeutic uses.

Objective and methodology

  • The key objective of the study was to discern the temporal changes in gene and protein expression during the establishment of equine MPC cultures.
  • The researchers obtained bone marrow aspirate from 35 horses and processed the samples using density gradient centrifugation. Subsequently, they studied the gene and protein expression at 2, 7, 14, 21, and 30 days of cell culture.

Findings and implications

  • Initially, bone marrow mononuclear cells showed variable expression of CD44, CD11a/CD18, CD90, and CD45RB surface molecules. But two hours post culture, the cells displayed a specific phenotype of active CD44 and CD29, with low CD90 and CD45RB expression.
  • There was good agreement in gene and protein expression in cell cultures across all the time-points studied, showing the gene-protein expression consistency of the cells during cell culture.
  • In established MPC cultures, cells remained strongly positive for CD44 and CD29. This implies that these molecules are steadfast markers of the established MPC cultures.
  • The intensity and proportion of cells expressing CD90 increased as the culture became more homogeneous. This could indicate an evolution of cell phenotype as the culture conditions stabilise over time.
  • Conversely, CD11a/CD18 and CD45RB expression decreased over time. This is significant as this decrease can act as suitable negative markers for MPC cultures under ordinary growth conditions.
  • The study appears to be first of its kind to show dynamic changes in cell surface molecule expression during the early phase of MPC culture establishment. This can offer new insights into improving MPC isolation procedures in research or therapeutics setup.

Cite This Article

APA
Radcliffe CH, Flaminio MJ, Fortier LA. (2009). Temporal analysis of equine bone marrow aspirate during establishment of putative mesenchymal progenitor cell populations. Stem Cells Dev, 19(2), 269-282. https://doi.org/10.1089/scd.2009.0091

Publication

Stem cells and development
ISSN: 1557-8534
NlmUniqueID: 101197107
Country: United States
Language: English
Volume: 19
Issue: 2
Pages: 269-282

Researcher Affiliations

Radcliffe, Catherine H
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York.
Flaminio, M Julia B F
    Fortier, Lisa A

      MeSH Terms

      • Animals
      • Bone Marrow Cells / cytology
      • Bone Marrow Cells / metabolism
      • Cell Cycle
      • Cell Differentiation
      • Cell Separation / methods
      • Cells, Cultured
      • Flow Cytometry
      • Gene Expression Profiling
      • Horses
      • Hyaluronan Receptors / genetics
      • Hyaluronan Receptors / metabolism
      • Immunophenotyping
      • Integrin beta1 / genetics
      • Integrin beta1 / metabolism
      • Leukocytes, Mononuclear / cytology
      • Leukocytes, Mononuclear / metabolism
      • Mesenchymal Stem Cells / cytology
      • Mesenchymal Stem Cells / metabolism
      • Molecular Sequence Data
      • Reverse Transcriptase Polymerase Chain Reaction
      • Time Factors

      Grant Funding

      • T32 RR007059 / NCRR NIH HHS
      • T32RR07059 / NCRR NIH HHS

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