3D Bioprinting Human Induced Pluripotent Stem Cell-Derived Neural Tissues Using a Novel Lab-on-a-Printer Technology

Abstract

Most neurological diseases and disorders lack true cures, including spinal cord injury (SCI). Accordingly, current treatments only alleviate the symptoms of these neurological diseases and disorders. Engineered neural tissues derived from human induced pluripotent stem cells (hiPSCs) can serve as powerful tools to identify drug targets for treating such diseases and disorders. In this work, we demonstrate how hiPSC-derived neural progenitor cells (NPCs) can be bioprinted into defined structures using Aspect Biosystems’ novel RX1 bioprinter in combination with our unique fibrin-based bioink in rapid fashion as it takes under 5 min to print four tissues. This printing process preserves high levels of cell viability (>81%) and their differentiation capacity in comparison to less sophisticated bioprinting methods. These bioprinted neural tissues expressed the neuronal marker, βT-III (45 ± 20.9%), after 15 days of culture and markers associated with spinal cord (SC) motor neurons (MNs), such as Olig2 (68.8 ± 6.9%), and HB9 (99.6 ± 0.4%) as indicated by flow cytometry. The bioprinted neural tissues expressed the mature MN marker, ChaT, after 30 days of culture as indicated by immunocytochemistry. In conclusion, we have presented a novel method for high throughput production of mature hiPSC-derived neural tissues with defined structures that resemble those found in the SC.

3D Bioprinting Pluripotent Stem Cell Derived Neural Tissues Using a Novel Fibrin Bioink Containing Drug Releasing Microspheres

“…Such bioprinted microenvironments can promote the differentiation of hiPSC-derived NPCs into mature, electrophysiologically active neurons. Our group has engineered 3D bioprinted hiPSC-derived neural tissue that mimics spinal cord tissue by treating these tissues with a variety of small molecules (De La Vega et al, 2018a). While these 3D bioprinted constructs show promise as an in vitro neural tissue models, there is still significant room for improvement.…”

“…Our own group developed a novel fibrin-based bioink for printing hiPSC-derived neural aggregates that both maintained their viability and differentiated into mature neural tissues after 46 days of culture (Abelseth et al, 2018). This same formulation was also used to print dissociated hiPSC-derived neural progenitor cells (NPCs) that could be matured into spinal cord-resembling tissues upon treatment with specific small molecules (De La Vega et al, 2018a). This bioink formulation supported the generation of ring shaped constructs containing the human glioblastoma cell line where the tissues exhibited high levels of viability and expressed cancer associated protein markers (Lee et al, 2019).…”

3D Printed Neural Regeneration Devices

“…However, constructing multicellular architectures remains a significant challenge. Recently, commercially available extrusion‐based lab‐on‐a‐printer systems have been utilized for tissue engineering studies . These printers consist of a variable system which can control the desired temperature inside the syringe, a printing stage which is tunable in the range of 4–37 °C depending on the specific requirement of the cell‐hydrogel suspension for maintain cell viability, and a built‐in ultraviolet (UV) light system for sterility and cross‐linking.…”

“…Moreover, depending on the specific requirement for the printing process, the printer can be customized. For instance, to reduce shear stress on cells during extrusion, Willerth et al have integrated microfluidic channels into the printhead, which allowed a separate flow of cells in bioinks and the associated cross‐linker . This enabled low viscous flow, resulting in successful neural differentiation from printed human induced pluripotent stem cells (iPSCs).…”

Engineered 3D Polymer and Hydrogel Microenvironments for Cell Culture Applications

“…Fibrin hydrogel encapsulated bone marrow stromal cells were inkjet printed and showed cytocompatibility of the encapsulation material [307]. Fibrin has been used as the base bioink for the printing of neural tissue as a glioblastoma model for drug screening [308]; human dental pulp stem cells for tooth tissue engineering [309]; Schwann cells for nerve tissue engineering [219,303]; human umbilical vein endothelial cells, and hMSCs for bone tissue engineering and neovascularization [310]; as well as human induced pluripotent stem cells for neurological diseases [311].…”