Abstract
Previously, we showed that mesenchymal stem cells (MSC) can be mobilized into peripheral blood using electroacupuncture (EA) at acupoints, LI-4, LI-11, GV-14, and GV-20. The purpose of this study was to determine whether EA-mobilized MSC could be harvested and expanded in vitro to be used as an autologous cell therapy in horses. Peripheral blood mononuclear cells (PBMC) isolated from young and aged lame horses (n = 29) showed a marked enrichment for MSCs. MSC were expanded in vitro (n = 25) and administered intravenously at a dose of 50 x 106 (n = 24). Treatment resulted in significant improvement in lameness as assessed by the American Association of Equine Practitioners (AAEP) lameness scale (n = 23). MSCs exhibited immunomodulatory function by inhibition of lymphocyte proliferation and induction of IL-10. Intradermal testing showed no immediate or delayed immune reactions to MSC (1 x 106 to 1 x 104). In this study, we demonstrated an efficient, safe and reproducible method to mobilize and expand, in vitro, MSCs in sufficiently high concentrations for therapeutic administration. We confirm the immunomodulatory function of these cells in vitro. This non-pharmacological and non-surgical strategy for stem cell harvest has a broad range of biomedical applications and represents an improved clinically translatable and economical cell source for humans.
Highlights
Mesenchymal stem cells (MSC) are predominately derived from adipose tissue (AT) and bone marrow (BM) and have beneficial effects in a variety of clinical conditions [1, 2] including pain relief, immune modulation and tissue regeneration [3]
This supports the feasibility of using Horse Recombinant Medium (HRM) +15% autologous horse serum for expansion of equine MSC and for minimizing allergic reactions with repeated administration. This was confirmed in two horses that received autologous MSC expanded with HRM+15% autologous serum and did not experience any adverse reaction. This is the first report of peripheral blood-derived MSC for therapeutic acquisition without the use of drugs or growth factors for mobilization
Advanced age did not appear to be a limitation to the mobilization and expansion of EA mobilized MSC (EA-MSC) as these cells were isolated from elderly horses (S1 Table), and the cells maintained a robust growth rate in culture allowing clinical application in a population of horses most in need of treatment
Summary
Mesenchymal stem cells (MSC) are predominately derived from adipose tissue (AT) and bone marrow (BM) and have beneficial effects in a variety of clinical conditions [1, 2] including pain relief, immune modulation and tissue regeneration [3]. Bone marrow-derived MSC (BM-MSC) have been shown to result in improved tissue repair in experimental models of equine tendinopathy as well as in naturally occurring disease [4,5,6]. Minimally invasive AT collection in the form of liposuction is not widely available. Prior attempts to isolate MSC from equine blood without mobilization as a simple and minimally invasive source of MSC were unsuccessful [10]
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