Hyunjin Yoo1, Dongyoon Choi1, Youngsok Choi1. 1. Department of Stem Cell and Regenerative Biotechnology, Institute of Advanced and Regenerative Science, Konkuk University, Seoul 05029, Korea.
Abstract
OBJECTIVE: Pluripotent 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]). METHODS: We 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. RESULTS: We 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. CONCLUSION: Together, 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.
OBJECTIVE: Pluripotent 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]). METHODS: We 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. RESULTS: We 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. CONCLUSION: Together, 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.
Authors: Marika J Karkkainen; Paula Haiko; Kirsi Sainio; Juha Partanen; Jussi Taipale; Tatiana V Petrova; Michael Jeltsch; David G Jackson; Marja Talikka; Heikki Rauvala; Christer Betsholtz; Kari Alitalo Journal: Nat Immunol Date: 2003-11-23 Impact factor: 25.606
Authors: Mathias François; Andrea Caprini; Brett Hosking; Fabrizio Orsenigo; Dagmar Wilhelm; Catherine Browne; Karri Paavonen; Tara Karnezis; Ramin Shayan; Meredith Downes; Tara Davidson; Desmond Tutt; Kathryn S E Cheah; Steven A Stacker; George E O Muscat; Marc G Achen; Elisabetta Dejana; Peter Koopman Journal: Nature Date: 2008-10-19 Impact factor: 49.962