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Updated: Mar 25, 2026
Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits
Published on: June 2, 2019
Expansion and angiogenic potential of mesenchymal stem cells from patients with critical limb ischemia
Luke Brewster1, Scott Robinson2, Ruoya Wang2
1Department of Surgery, Emory University School of Medicine, Atlanta, Ga; Surgical and Research Services, Atlanta Veterans Affairs Medical Center, Atlanta, Ga; Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Ga.
Insights
Platelet lysate (PL) significantly enhances the expansion of mesenchymal stem cells (MSCs) from critical limb ischemia (CLI) patients. These expanded MSCs retain their angiogenic potential, offering a promising cell therapy for CLI.
Area of Science:
- Regenerative Medicine
- Cellular Therapy
- Vascular Biology
Background:
- Critical limb ischemia (CLI) is a severe condition with limited treatment options, where cellular therapies like mesenchymal stem cells (MSCs) show promise.
- Challenges in MSC therapy for CLI include poor cell quality and inadequate post-delivery viability.
- Autologous MSCs from CLI patients may have impaired expansion and angiogenic potential.
Purpose of the Study:
- To evaluate the efficacy of pooled human platelet lysate (PL) as a culture supplement for expanding MSCs from CLI patients.
- To compare the in vitro angiogenic activity of MSCs expanded with PL versus standard fetal bovine serum (FBS).
- To assess the angiogenic potential of MSCs derived from CLI patients compared to healthy donors.
Main Methods:
- Mesenchymal stem cells (MSCs) were isolated from CLI patients (with and without diabetes) and healthy donors.
- MSCs were cultured using either PL or FBS as a supplement.
- Phenotypic markers, differentiation capacity, population doublings, and in vitro angiogenic activity were assessed.
Main Results:
- Platelet lysate (PL) significantly increased the population doublings of MSCs from all donor groups compared to FBS.
- MSCs from CLI patients exhibited angiogenic effects on endothelial cells comparable to those from healthy donors.
- PL supplementation did not negatively impact the angiogenic activity of MSCs.
Conclusions:
- Pooled human platelet lysate (PL) effectively promotes the rapid expansion of MSCs from both CLI patients and healthy individuals.
- MSCs expanded from CLI patients demonstrate significant angiogenic activity, comparable to healthy counterparts.
- PL-expanded autologous MSCs offer a viable strategy for improving cell therapy delivery and efficacy in CLI patients.
Background:
Critical limb ischemia (CLI) is a life- and limb-threatening condition affecting 1% to 10% of the population with peripheral arterial disease. Traditional revascularization options are not possible for up to 50% of CLI patients, in which case, the use of cellular therapies, such as bone marrow-derived mesenchymal stem cells (MSCs), hold great promise as an alternative revascularization therapy. However, no randomized, controlled phase 3 trials to date have demonstrated an improvement in limb salvage with cellular therapies. This may be due to poor cell quality (ie, inability to generate a sufficient number of angiogenic MSCs) or to the inadequate retention and viability of MSCs after delivery, or both. Because concerns remain about the expansion and angiogenic potential of autologous MSCs in the CLI population, the objective of this study was to examine the effect of our novel culture media supplement, pooled human platelet lysate (PL), in lieu of the standard fetal bovine serum (FBS), to improve the expansion potential of MSCs from CLI patients. We also characterized the in vitro angiogenic activity of MSCs from the tibia of amputated CLI limbs compared with MSCs from healthy donors.
Methods:
MSCs were obtained from the tibia of four CLI patients (ISC) and four ISC patients with diabetes mellitus (ISC+DM) undergoing major amputation. Healthy MSCs were aspirated from the iliac crest of four young and healthy donors. MSCs were isolated and expanded in culture with PL or FBS. MSCs from passage 3 to 6 were used for phenotypic marker expression and for adipogenic and osteogenic differentiation and were tested for their in vitro angiogenic activity on human microdermal endothelial cells. In parallel MSCs were cultured to passage 11 for population-doubling calculations.
Results:
MSCs from ISC and ISC+DM patients and from healthy patients exhibited appropriate expression of cell surface markers and differentiation capacity. Population doublings were significantly greater for PL-stimulated compared with FBS-stimulated MSCs in all groups. Biologically active amounts of angiogens were identified in the secretome of all MSCs without consistent trends among groups. PL expansion did not adversely affect the angiogenic activity of MSCs compared with FBS. The ISC and ISC+DM MSCs demonstrated angiogenic effects on endothelial cells similar to those of healthy and ISC MSCs.
Conclusions:
PL promotes the rapid expansion of MSCs from CLI and healthy persons. Importantly, MSCs expanded from CLI patients demonstrate the desired angiogenic activity compared with their healthy counterparts. We conclude that autologous MSCs from CLI patients can be sufficiently expanded with PL and be expected to deliver requisite angiogenic effects in vivo. We expect the improved expansion of ISC and ISC+DM with PL to be helpful in improving the successful delivery of autologous MSCs to patients with CLI.
