Gene expression profiling of progenitor cells isolated from rat rotator cuff musculotendinous junction

BMC Musculoskelet Disord. 2020 Mar 28;21(1):194. doi: 10.1186/s12891-020-03190-9.

Abstract

Background: Rotator cuff tendon tears are typically degenerative and usually affect the region of tendon insertion on bone. The remnant torn tendon is degenerative and may not be an ideal source for progenitor cells for cell-based therapies. Therefore, the aim of this study was to determine if musculotendinous junction (MTJ), which is adjacent to tendon would be a viable alternate source of progenitor stem cells. We also sought to study the gene expression profile MTJ progenitors and compare it with progenitors isolated from RC tendon, RC muscle and other existing tissue sources (bone marrow, adipose tissue, and Achilles tendon).

Methods: Rotator cuff tendon (RCT), muscle (RCM), and RCMTJ as well as Achilles tendon (AT) tissues were harvested from healthy male Lewis rats and progenitor cultures were established from these tissues and also from bone marrow and adipose tissue. Quantitative RT-PCR was performed on RNA extracts from intact tissues and progenitor cells using a custom array for the mesenchymal stem cell (MSC) differentiation marker genes. The gene expression profile of MSC differentiation markers within four tissues types, six progenitor cells, and between tissue and their corresponding progenitors were compared.

Results: Progenitors cells can be isolated from rat rotator cuff musculotendinous tissue and their pattern of MSC gene expression was similar to the rotator cuff tendon progenitors for majority of the genes tested. However, there were significant differences between the MSC gene expression patterns of RCMTJ and RCM progenitors. Furthermore, there were differences in gene expression between the RCMTJ tissue and its progenitor cells with respect to MSC differentiation markers. The gene expression pattern of RCMTJ tissue was similar to RCM tissue with respect to markers of chondrogenesis, myogenesis, tenogenesis, and MSC specific markers.

Conclusion: We demonstrate that the musculotendinous junction contains distinct set of progenitor cells and their MSC gene expression pattern is similar to rotator cuff tendon progenitors. RCMTJ progenitors will be an attractive option for cell-based regenerative treatment of chronic rotator cuff tears.

Keywords: Bone marrow cells; MSC gene expression profile; Progenitor cell; Rotator cuff muscle; Rotator cuff musculotendinous junction; Rotator cuff tendon.

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Separation / methods*
  • Cells, Cultured
  • Chondrogenesis / genetics*
  • Feasibility Studies
  • Gene Expression Profiling
  • Humans
  • Male
  • Primary Cell Culture
  • Rats
  • Rotator Cuff / cytology*
  • Rotator Cuff Injuries / therapy
  • Stem Cell Transplantation
  • Stem Cells / metabolism*
  • Wound Healing / genetics

Substances

  • Biomarkers