Multipotent Mesenchymal Stem Cells Acquire a Lymphendothelial Phenotype and Enhance Lymphatic Regeneration In Vivo

Abstract

Background-The importance and therapeutic value of stem cells in lymphangiogenesis are poorly understood. We evaluated the potential of human and murine mesenchymal stem cells (MSCs) to acquire a lymphatic phenotype in vitro and to enhance lymphatic regeneration in vivo. Methods and Results-We assessed the lymphendothelial differentiation of human and murine MSCs after induction with supernatant derived from human dermal microvascular endothelial cells, isolated lymphatic endothelial cells, and purified vascular endothelial growth factor (VEGF)-C in vitro. We used human or murine progenitor MSC lines and then characterized the lymphatic phenotype by morphology, migratory capacity, and the expression of lymphatic markers such as Prox-1, podoplanin, Lyve-1, VEGF receptor-2, and VEGF receptor-3. Using a murine lymphatic edema model, we assessed the potential of these cells to form a functional lymphatic vasculature in vivo after injection of syngeneic MSCs. Incubation with supernatant from lymphatic endothelial cells induced an endothelium-like morphology and the expression of lymphendothelial markers in both human and murine MSCs in vitro. MSCs showed migratory activity along a VEGF-C gradient, which was enhanced by VEGF-C conditioning. In vivo, the local application of MSCs resulted in a significant decrease in edema formation (Ϫ20.1%; PϽ0.01 versus untreated tails) after 3 weekly cell injections and restored the drainage of intradermally injected methylene blue after 7 weekly injections. Conclusions-MSCs

Lymphatic endothelial progenitors originate from plastic myeloid cells activated by toll-like receptor-4

“…Although LECP were reported to derive from adipose [31], mesenchymal [30] or hematopoietic [59] stem cells, the majority of studies identified BM-produced myeloid-monocytic precursors as their main source [26]. Consistent with the monocytic origin, human LECP were successfully differentiated in vitro using monocytes from peripheral [27,32,33] or umbilical cord [24,54] blood and treatment with a VEGF-A/VEGF-C cocktail.…”

“…Such pro-lymphatic reprogramming has been shown for human monocytes isolated from peripheral or cord blood [24,27], human pluripotent stem cell lines [25], mouse embryonic cells [28], mouse BM-derived CD11b + and mononuclear cells [13,16,29], mouse and human mesenchymal stem cells [30] and adipose-derived stem cells [31]. The main criteria for defining differentiated cells as LECP are as follows: 1) expression of specific LEC markers [16,24,25,27]; 2) acquisition of an endothelial-specific cobblestone morphology and/or ability to form tubes when grown in matrigel [16]; 3) demonstrated function in vivo evidenced by integration into lymphatic vessels [12,15,22] and a statistically significant increase in lymphatic vessel density (LVD) in inflammatory and tissue remodeling models [24,25,32]; and 4) evidence for enhanced functionality of new lymphatics such as improved relief from lymphedema [32] and an accelerated rate of healing wounds [25].…”

Bone Marrow-Derived Mesenchymal Stem Cells Drive Lymphangiogenesis

“…In addition to acquire a lymphatic phenotype in vitro and induce lymphatic regeneration in vivo [48], MSC can directly take part to vessel formation by transdifferentiation into endothelial cells and incorporation into the vessel wall [4], [49]. Furthermore, they can secrete several factors implicated in angiogenesis such as VEGF-A, angiopoietin-1 and bFGF [20], [50].…”

Impact of the Prolymphangiogenic Crosstalk in the Tumor Microenvironment on Lymphatic Cancer Metastasis

“…MSCs are able to express a lymphatic phenotype, when cultured in lymphatic induced medium and VEGF-C [115]. Vice versa, tumor cells secrete growth factors, cytokines, and chemokines to promote the migration and survival of MSCs [108].…”