In VitroInduction of Human Adipose-Derived Stem Cells into Lymphatic Endothelial-Like Cells

Abstract

Human adipose-derived stem cells (hADSCs) may provide a suitable number of progenitors for the treatment of lymphatic edema; however, to date the protocols for inducing hADSCs into this tissue type have not been standardized. We wished to investigate the induction of hADSCs into lymphatic endothelial-like cells using vascular endothelial growth factor-C156S (VEGF-C156S) and other growth factors in vitro. hADSCs from healthy adult adipose tissue were purified using enzyme digestion. Differentiation was induced using medium containing VEGF-C156S and bovine fibroblast growth factor (bFGF). Differentiation was confirmed using immunostaining for lymphatic vessel endothelial hyaluronan receptor (LYVE-1) and fms-related tyrosine kinase 4 (FLT-4), two lymphatic endothelial cell markers. The expression levels of LYVE-1, prospero homeobox 1 (PROX-1), and FLT-4 throughout induction were assessed using reverse transcriptase quantitative polymerase chain reaction. hADSCs were successfully obtained by trypsin digest and purification. Flow cytometry showed these cells were similar to mesenchymal stem cells, with a high positive rate of CD13, CD29, CD44, and CD105, and a low positive rate of CD31, CD34, CD45, and HLA-DR. Induction to lymphatic endothelial-like cells was successful, with cells expressing high levels of LYVE-1, PROX-1, and FLT-4. Adipose-derived stem cells can be induced to differentiate into lymphatic endothelial-like cells using a medium containing VEGF-C156S, bFGF, and other growth factors. This population of lymphatic endothelial-like cells may be useful for lymphatic reconstruction in the future.

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].…”

Ginsenoside Rg1 and platelet-rich fibrin enhance human breast adipose-derived stem cells function for soft tissue regeneration

“…Yang reported that HBASCs have the potential to transdifferentiate into human mammary epithelial-like cells when co-cultured with breast epithelial cells (HBL-100 cell line) [34]. Das indicated that HBASCs induce penile angiogenesis and neural regeneration without systemic inflammation in diabetic mice [35].…”

IL‐7 enhances the differentiation of adipose‐derived stem cells toward lymphatic endothelial cells through AKT signaling

“…However, these methods are not only complicated but are accompanied by potential safety problem. Yang et al (2015) also found combination of VEGF-C C156s and bFGF can induce Human Adipose-Derived Stem Cells into Lymphatic Endothelial-Like Cells. It was reported that IL-7 maintained expression of Prox-1 and Podoplanin in blood endothelial cells (BECs), increased expression of lymphatic markers LYVE-1, and Podoplanin, and stimulated tubulogenesis and capillary sprouting (Al-Rawi et al, 2005a).…”

“…Specific gene based reprogramming is one of the methods to alter the cellular fate of cells (Deng et al, 2017). To test our hypothesis, IL-7 was added to the traditional induced medium including VEGF-A, VEGF-C, and FGF (Conrad et al, 2009;Lee et al, 2010Lee et al, , 2015Takeda et al, 2015;Yang et al, 2015). However, these methods are not only complicated but are accompanied by potential safety problem.…”