Behavior of CMPCs in unidirectional constrained and stress-free 3D hydrogels.
Cardiomyocyte progenitor cells (CMPCs) are a candidate cell source for cardiac regenerative therapy. However, like other stem cells, after transplantation in the heart, cell retention and differentiation capacity of the CMPCs are low. Combining cells with biomaterials might overcome this problem. By serving as a (temporal) environment, the biomaterial can retain the cells and provide signals that enhance survival, proliferation and differentiation of the cells. To gain more insight into the effect that the encapsulation of CMPCs in a biomaterial has on their behavior, we cultured CMPCs in unidirectional constrained and stress-free collagen/Matrigel hydrogels. CMPCs cultured in 3D hydrogels stay viable and keep their cardiomyogenic profile independent of the application of strain. Moreover, the increased expression of Nkx2.5, myocardin and cTnT in 3D hydrogels compared to 2D cultures, suggests enhanced cardiomyogenic differentiation capacity of cells in 3D. Furthermore, increased expression of collagen I, collagen III, elastin and fibronectin and of the matrix remodeling enzymes MMP-1, MMP-2, MMP-9, and TIMP-1 and TIMP-2 in the 3D hydrogels is indicative of an enhanced matrix remodeling capacity of CMPCs in a 3D environment, independent of the application of strain. Interestingly, the additional application of static strain to the 3D hydrogels, as imposed by hydrogel constrainment, stabilized CMPC viability and proliferation, resulted in enhanced cardiac marker protein expression and appeared crucial for cellular organization and morphology. More specifically, CMPCs cultured in 3D collagen/Matrigel constrained hydrogels became readily mechanosensitive, had a rod-shaped morphology, and responded to the applied strain by orienting in the direction of the constraint. Overall, our data demonstrate the applicability of CMPCs in a 3D environment since encapsulation of CMPCs may stabilize survival and proliferation, can enhance the differentiation and remodeling capacity of the cells, and could induce cellular re-organization, which all may contribute to an improved efficiency of cardiac stem cell therapy.
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: m.h.v.marion@tue.nl.
Mieke H
MH
van Marion
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands; Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: n.a.m.bax@tue.nl.
Noortje A M
NA
Bax
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: m.c.v.turnhout@tue.nl.
Mark C
MC
van Turnhout
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: a.mauretti@tue.nl.
Arianna
A
Mauretti
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: d.w.j.v.d.schaft@tue.nl.
Daisy W J
DW
van der Schaft
Department of Molecular Cell Biology and Center for Biomedical Genetics, Leiden University Medical Center, The Netherlands. Electronic address: M.J.T.H.Goumans@lumc.nl.
Marie José T H
MJ
Goumans
Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands; Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: c.v.c.bouten@tue.nl.
Carlijn V C
CV
Bouten
D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D005353:Fibronectins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D020782:Matrix Metalloproteinases / Q000096:biosynthesis; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000637:transplantation*; D011509:Proteoglycans / Q000494:pharmacology; D044968:Regenerative Medicine; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis
D001672:Biocompatible Materials; D004338:Drug Combinations; D005353:Fibronectins; D007797:Laminin; D011509:Proteoglycans; C560822:TIMP1 protein, human; C560823:TIMP2 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; C059485:matrigel; D019716:Tissue Inhibitor of Metalloproteinase-2; D003094:Collagen; D020782:Matrix Metalloproteinases
3D cell culture; Cardiac regeneration; Cardiomyocyte progenitor cells (CMPCs); Matrix remodeling; Mechanical forces; Tissue engineering