Long-term Culture of Human iPS Cell-derived Telencephalic Neuron Aggregates on Collagen Gel

Abstract

It takes several months to form the 3-dimensional morphology of the human embryonic brain. Therefore, establishing a long-term culture method for neuronal tissues derived from human induced pluripotent stem (iPS) cells is very important for studying human brain development. However, it is difficult to keep primary neurons alive for more than 3 weeks in culture. Moreover, long-term adherent culture to maintain the morphology of telencephalic neuron aggregates induced from human iPS cells is also difficult. Although collagen gel has been widely used to support long-term culture of cells, it is not clear whether human iPS cell-derived neuron aggregates can be cultured for long periods on this substrate. In the present study, we differentiated human iPS cells to telencephalic neuron aggregates and examined long-term culture of these aggregates on collagen gel. The results indicated that these aggregates could be cultured for over 3 months by adhering tightly onto collagen gel. Furthermore, telencephalic neuronal precursors within these aggregates matured over time and formed layered structures. Thus, long-term culture of telencephalic neuron aggregates derived from human iPS cells on collagen gel would be useful for studying human cerebral cortex development.Key words: Induced pluripotent stem cell, forebrain neuron, collagen gel, long-term culture.

Differential Effects of Heparin and Hyaluronic Acid on Neural Patterning of Human Induced Pluripotent Stem Cells

“…During brain organoid formation, extracellular matrix (ECM) hydrogels, − such as Matrigel, collagen gels, and poly­(ethylene glycol) (PEG) hydrogels, − can provide instructive extracellular signals for hiPSC self-assembly into aggregates and facilitate neural tissue development . Matrigel is an undefined extract derived from mouse Engelbreth–Holm–Swarm tumors and composed of 60% laminin.…”

“…5,11 Therefore, the spatially compartmentalized microenvironments and the spatial-temporal regulation of the induction factor gradients become important in developing the next generation of brain organoids. 11,12 During brain organoid formation, extracellular matrix (ECM) hydrogels, 11−15 such as Matrigel, 1 collagen gels, 16 and poly(ethylene glycol) (PEG) hydrogels, 17−19 can provide instructive extracellular signals for hiPSC self-assembly into aggregates and facilitate neural tissue development. 8 Matrigel is an undefined extract derived from mouse Engelbreth−Holm− Swarm tumors and composed of 60% laminin.…”

Lab-on-Chip Microsystems for Ex Vivo Network of Neurons Studies: A Review

“…The microenvironment decides the direction of cell differentiation, cellular vitality, and lifetime. Plenty of methods have been proposed for long-term cell culture and maintaining a stable extracellular microenvironment ( Choi et al, 2015 ; Oyama et al, 2018 ; Mun et al, 2020 ). Scientists may change the extracellular environment deliberately to build up a certain disease model in order to show how the changes in the microenvironment in the brain cause the onset of a disease ( Osaki et al, 2018 ).…”

Microfluidic Particle Reactors: From Interface Characteristics to Cells and Drugs Related Biomedical Applications

“…The DCP microparticles can be employed to simulate natural red blood cells and cultivate human mesenchymal stem cells impacted by their remarkable interface characteristics (such as softness to pass through narrow channels and cytocompatibility toward human stem cells) ( Figure a). [ 85 ] 2) Collagen or fibrin has a unique surface topology and cell adhesion peptides, [ 86–90 ] allowing for the guidance of a series of cell behaviors (such as cell adhesion, proliferation, and differentiation) (Figure 2b). [ 91 ] Collagen generally requires thermal gelation while fibrin gel needs an enzymatic cross‐linking reaction with thrombin to gelation.…”

“…Collagen or fibrin has a unique surface topology and cell adhesion peptides, [86][87][88][89][90] allowing for the guidance of a series of cell behaviors (such as cell adhesion, proliferation, and differentiation) (Figure 2b). [91] Collagen generally requires thermal gelation while fibrin gel needs an enzymatic cross-linking reaction with thrombin to gelation.…”