Laser cell-micropatterned pair of cardiomyocytes: the relationship between basement membrane development and gap junction maturation

Abstract

The basement membrane (BM), a network of laminin and collagen IV, mechanically supports individual cells and directly mediates cell-cell and cell-extracellular matrix (ECM) interactions. For example, the BM network that tightly encloses each cardiomyocyte (CM) mediates the alignment of CMs with collagen I in the ECM. Additionally, the BM-laminin is involved in the formation of gap junctions (GJs), which regulate electrical coupling between two CMs in the myocardium. The role of BM in GJ maturation remains unclear because of the complicated in vivo structures and lack of an ideal in vitro culturing mode. In this study, our laser cell-micropatterning system was used to place two neonatal CMs (NCMs) in contact on an aligned collagen gel (ACG) to study the relationship between GJ maturation and BM development. The results of double immunofluorescence staining and confocal imaging showed that BM-laminin was deposited earlier than the formation of GJs in the intercellular space and that newly expressed connexin 43 clusters were preferentially assembled near the deposited BM structures. Eventually the BM network surrounded the GJs.

Dynamic Myofibrillar Remodeling in Live Cardiomyocytes under Static Stretch

“…Neonatal ventricular CMs were harvested from the rats using a two-day protocol. The detailed procedure is described in our previous publications 38 39 .…”

Embryo implantation triggers dynamic spatiotemporal expression of the basement membrane toolkit during uterine reprogramming

“…It is known that cell-cell junctions are features of decidua establishment and function as macromolecular barriers between the mother and embryo which can block penetrance of IgG that can elicit unwanted immune rejection of the embryo and add rigidity to the tissue [86,87]. Interestingly, a recent study on cardiomyocytes, a mesenchymal-derived cell encapsulated by BM, showed a similar pericellular BM that underwent alterations for the mediation of cell junction formation [88]. Perhaps decidua BM also mediates tight junction formation and functions as an additional selective barrier and contributes to rigidly while junctions are established.…”

Current methods for fabricating 3D cardiac engineered constructs

“…One such is the use of laser tweezers for single-cell micropatterning. A study utilized this technology to prepare the pattern and identify interactions between CMs, specifically at their gap junctions ( Yang et al., 2014a ). These laser etching techniques offer the unique ability to fabricate cardiac tissue structure with high degrees of resolution both consistently and efficiently through the vaporization of the material ( Gittard and Narayan, 2014 ).…”

“…They contain sarcolemma for anchoring the integrin and receptor transmembrane proteins that bind the CM to the extracellular matrix (ECM), including cardiac basement membrane ( Allyson Walker and Francis Spinale, 1999 ; Yang et al., 2015 ). The cardiac basement membrane plays key roles in sarcomerogenesis, CM-CM interaction, and cardiac electrophysiology based on the findings in our previous engineering models ( Yang et al., 2014b ; Yang et al., 2014 ; Yang et al., 2015 ). The sarcoplasmic reticulum in CMs stores the Ca 2+ used in the excitation-contraction coupling ( Williams, 1997 ).…”