Abstract
Herein, we report the fabrication of native organ-like three-dimensional (3D) cardiac tissue with an oriented structure and vascular network using a layer-by-layer (LbL), cell accumulation and 3D printing technique for regenerative medicine and pharmaceutical applications. We firstly evaluated the 3D shaping ability of hydroxybutyl chitosan (HBC), a thermoresponsive polymer, by using a robotic dispensing 3D printer. Next, we tried to fabricate orientation-controlled 3D cardiac tissue using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and normal human cardiac fibroblasts (NHCF) coated with extracellular matrix (ECM) nanofilms by layer-by-layer technique. These cells were seeded in the fabricated rectangular shape HBC gel frame. After cultivation of the fabricated tissue, fluorescence staining of the cytoskeleton revealed that hiPSC-CM and NHCF were aligned in one direction. Moreover, we were able to measure its contractile behavior using a video image analysis system. These results indicate that orientation-controlled cardiac tissue has more remarkable contractile function than uncontrolled cardiac tissue. Finally, co-culture with human cardiac microvascular endothelial cells (HMVEC) successfully provided a vascular network in orientation-controlled 3D cardiac tissue. The constructed 3D cardiac tissue with an oriented structure and vascular network would be a useful tool for regenerative medicine and pharmaceutical applications.
Highlights
Tissue engineering technology uses living cells, cytokines and biomaterials with the primary goal of creating functional tissue substitutes for novel medical treatments and pharmaceutical development[1]
Assessment of hydroxybutyl chitosan (HBC) gel printing by robotic dispensing 3D printer
Artificial 3D tissues are in great demand in regenerative medicine and the drug development field for medical treatments and in vitro assays
Summary
Tissue engineering technology uses living cells, cytokines and biomaterials with the primary goal of creating functional tissue substitutes for novel medical treatments and pharmaceutical development[1]. For the development of novel cell transplantation therapy and drug screening in vivo, it is important to fabricate cardiac tissue with the heart-specific structure of orientation in order to complement organ function. We report on a method for the fabrication of 3D cardiac tissue with heart specific structure, cell orientation and vascular network. To achieve this purpose, we reported on a fabrication method of orientation-controlled 3D tissue by using an LbL technique, cell accumulation method and 3D print technology[48]. We fabricated ECM nanofilms onto hiPSC-CM and normal human cardiac fibroblast (NHCF) cells by using the LbL technique These cells were seeded in the HBC gel frame by using the cell accumulation method to make orientation-controlled 3D tissue. We tried to fabricate native-like 3D cardiac tissue with orientation and vascular network constructs using co-cultured hiPSC-CM, NHCF and human cardiac microvascular endothelial cells (HMVEC)
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