Modeling the adhesion of human embryonic stem cells to poly(lactic-co-glycolic acid) surfaces in a 3D environment.
Human embryonic stem cells (hESCs) have previously been cultured on three dimensional (3D) biodegradable polymer scaffolds. Although complex structures were formed from the hESCs, very little is known about the mechanism of adhesion of these cells to the surfaces of the scaffolds. In this study, we achieved the efficient adhesion of pluripotent hESCs to 3D poly(lactic-co-glycolic acid) (PLGA) scaffolds based on our data from a novel two dimensional (2D) model that imitates the surface properties of the scaffolds. In the 2D model, single cell preparations of pluripotent hESCs adhered efficiently and predominantly to PLGA surfaces coated with laminin in comparison to collagen I, collagen IV, or fibronectin-coated surfaces. Flow cytometry analysis revealed that almost all of the pluripotent single cells expressed the integrin alpha 6, with a small percentage also expressing alpha 3ss1, which facilitates adhesion to laminin. This data was then translated into the 3D environment, with the efficient binding of single pluripotent hESCs to PLGA scaffolds coated with laminin. The utility of this system was shown by the directed differentiation of single hESCs seeded within laminin-coated scaffolds toward the endoderm lineage.
Gao
Steven Y
SY
Human Stem Cell Group, Diabetes Transplant Unit, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
Lees
Justin G
JG
Wong
Jennifer C Y
JC
Croll
Tristan I
TI
George
Peter
P
Cooper-White
Justin J
JJ
Tuch
Bernard E
BE
D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D053595:Embryonic Stem Cells / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology; D005353:Fibronectins / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D016023:Integrins / Q000737:chemistry / Q000378:metabolism; D019344:Lactic Acid / Q000737:chemistry*; D007797:Laminin / Q000378:metabolism; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013499:Surface Properties; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D018076:DNA, Complementary; D005353:Fibronectins; D016023:Integrins; D007797:Laminin; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D012313:RNA; D003094:Collagen