Conjugation of vascular endothelial growth factor to poly lactic-co-glycolic acid nanospheres enhances differentiation of embryonic stem cells to lymphatic endothelial cells.

Hyunjin Yoo,Dongyoon Choi,Youngsok Choi

doi:10.5713/ajas.20.0202

Hyunjin Yoo, Dongyoon Choi + Show 1 more

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https://doi.org/10.5713/ajas.20.0202

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Journal: Animal bioscience	Publication Date: Aug 21, 2020
License type: CC BY-NC 4.0

Affiliation: Konkuk University

Abstract

ObjectivePluripotent stem cell-derived lymphatic endothelial cells (LECs) show great promise in their therapeutic application in the field of regenerative medicine related to lymphatic vessels. We tested the approach of forced differentiation of mouse embryonal stem cells into LECs using biodegradable poly lactic-co-glycolic acid (PLGA) nanospheres in conjugation with growth factors (vascular endothelial growth factors [VEGF-A and VEGF-C]).MethodsWe evaluated the practical use of heparin-conjugated PLGA nanoparticles (molecular weight ~15,000) in conjugation with VEGF-A/C, embryoid body (EB) formation, and LEC differentiation using immunofluorescence staining followed by quantification and quantitative real-time polymerase chain reaction analysis.ResultsWe showed that formation and differentiation of EB with VEGF-A/C-conjugated PLGA nanospheres, compared to direct supplementation of VEGF-A/C to the EB differentiation media, greatly improved yield of LYVE1(+) LECs. Our analyses revealed that the enhanced potential of LEC differentiation using VEGF-A/C-conjugated PLGA nanospheres was mediated by elevation of expression of the genes that are important for lymphatic vessel formation.ConclusionTogether, we not only established an improved protocol for LEC differentiation using PLGA nanospheres but also provided a platform technology for the mechanistic study of LEC development in mammals.

Highlights

The lymphatic system plays essential roles in the host defense mechanism including the promotion of immune cell maturation, lipid reabsorption, and balance of interstitial fluid [1]
We analyzed the practical application of poly lactic-co-glycolic acid (PLGA) nanospheres in differentiation of lymphatic endothelial cells (LECs) from mouse embryonal stem cells and found that mouse embryoid body (EB) conjugated with vascular endothelial growth factor (VEGF)-A/C loaded heparin-conjugated-PLGA nanospheres enhanced LEC differentiation by enhancing the expression of the genes that are important for lymphangiogenesis
We determined a potential use of biodegradable PLGA nanospheres in conjugation with growth factors (GFs) (VEGFA and VEGF-C) and forced differentiation of mouse embryonal stem cells (mESCs) into LECs

Summary

Introduction

The lymphatic system plays essential roles in the host defense mechanism including the promotion of immune cell maturation, lipid reabsorption, and balance of interstitial fluid [1]. Tion as knockout of either VEGF-C or VEGFR3 in mouse models displayed embryonic lethality with aberrant lymphatic formation [6,7]. Consistent with this notion, the effects of VEGF-C/VEGFR3 led to the proliferation and migration of LECs and blood ECs (BECs) [8]. Previous studies demonstrated that LECs were successfully differentiated from both mouse and human pluripotent stem cells (PSCs) using VEGF-C/VEGFR3 signals [10,11]. A sustained release of fibronectin after conjugation with 3,4-dihydroxy-L-phenylalanine (L-DOPA)-coated PLGA improved in vitro neuronal differentiation of human mesenchymal stem cells. Our study established an improved protocol for LEC differentiation using PLGA nanospheres and provided a platform technology for the mechanistic study of LEC development in mammals

Methods

Results

Conclusion