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Derivation and Differentiation of Canine Ovarian Mesenchymal Stem Cells
Published on: December 16, 2018
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Immunophenotypic characterization of ovine mesenchymal stem cells
Mohammad R Khan1, Anil Chandrashekran1, Roger K W Smith1
1Clinical Sciences and Services, Royal Veterinary College, North Mymms, Hatfield, Herts, AL9 7TA, United Kingdom.
Summary
This study identified specific antibodies for characterizing sheep mesenchymal stem cells (MSCs). These validated tools are crucial for advancing stem cell therapies in large animal models relevant to human diseases.
Area of Science:
- Stem Cell Biology
- Immunology
- Animal Models
Background:
- Mesenchymal stem cells (MSCs) hold significant clinical potential.
- Characterizing MSCs requires specific analytical tools like antibodies.
- Existing tools are often unavailable for large animal models, such as sheep, hindering research into human musculoskeletal diseases.
Purpose of the Study:
- To identify commercially available, non-species-specific monoclonal antibodies for immunophenotypic characterization of ovine MSCs.
- To develop a reliable protocol for detaching ovine MSCs with minimal loss of cell surface antigens.
- To validate the cross-reactivity of these antibodies with ovine MSCs.
Main Methods:
- Developed a cell detachment protocol using accutase and EDTA.
- Tested a range of commercially available antibodies against human or rodent MSC antigens for cross-reactivity with ovine MSCs.
- Utilized single and multistain-based assays, including CD29, CD44, and CD166 as positive controls.
Main Results:
- Identified cross-reactive antibody clones for ovine CD73, CD90, CD105, CD271, and MHC1 antigens.
- Confirmed colocalization of these antigens on ovine MSCs via multistaining.
- Observed minimal to no cross-reactivity with ovine white blood cell antigens (CD117, MHCII, CD31, CD45).
Conclusions:
- Validated a panel of antibodies for immunophenotypic characterization of ovine MSCs.
- Established a reliable method for MSC detachment preserving cell surface markers.
- These findings are essential for utilizing sheep as a large animal model in stem cell-based therapies.
