Neural Stem Cell Differentiation Using Microfluidic Device-Generated Growth Factor Gradient

Abstract

Neural stem cells (NSCs) have the ability to self-renew and differentiate into multiple nervous system cell types. During embryonic development, the concentrations of soluble biological molecules have a critical role in controlling cell proliferation, migration, differentiation and apoptosis. In an effort to find optimal culture conditions for the generation of desired cell types in vitro, we used a microfluidic chip-generated growth factor gradient system. In the current study, NSCs in the microfluidic device remained healthy during the entire period of cell culture, and proliferated and differentiated in response to the concentration gradient of growth factors (epithermal growth factor and basic fibroblast growth factor). We also showed that overexpression of ASCL1 in NSCs increased neuronal differentiation depending on the concentration gradient of growth factors generated in the microfluidic gradient chip. The microfluidic system allowed us to study concentration-dependent effects of growth factors within a single device, while a traditional system requires multiple independent cultures using fixed growth factor concentrations. Our study suggests that the microfluidic gradient-generating chip is a powerful tool for determining the optimal culture conditions.

Regulation of Neural Stem Cell Fate by Natural Products

“…Thus, the protocols need to be optimized before they can be applied in regenerative medicine. Recently, in addition to soluble factors that are required for proper cell development, culture dish surfaces and microfluidic chambers that are effective in generating gradients of soluble factors have also been studied to identify the optimal environment for SC culture and differentiation (Cha et al ., 2017; Kim et al ., 2018).…”

Application of microfluidic systems for neural differentiation of cells

“…Results interestingly confirmed that the proliferation and differentiation of hNSCs were directly depended on GF concentration. [4,22] Many other bio-molecules can influence the differentiation of stem cells based on their physiologic role in specific organs in vivo.…”

“…Stem cells are widely considered as biopharmaceuticals and cell therapies due to their potential proliferation, genetic modification with external gene delivery systems, and differentiation to neurons, astrocytes, and oligodendrocytes. [4] Consequently, inducing the differentiation of stem cells toward neural cell lineages plays an important role in the repairing process. There are 3 main types of stem cells involved in trans-differentiation studies and cellular replacement therapy including embryonic stem cells (ESC), adult stem cells (ASC), and induced pluripotent stem cells (iPSC).…”

Biological Behavioral Alterations of the Post-neural Differentiated Dental Pulp Stem Cells Through an in situ Microenvironment

“…Recently, it has been confirmed that DPSCs can be induced to form neurospheres or differentiate into mature neurons by endogenous signaling cues ( Heng et al, 2016 ). Many factors, for instance, the extracellular matrix, mechanical force, growth factors, culture systems, oxygen stress, and microfluidic systems, have been reported as critical factors for the induction of neuronal differentiation ( Heng et al, 2016 ; Thompson and Chan, 2016 ; Yamazaki et al, 2016 ; Kim et al, 2018 ; Liu et al, 2020 ). However, few in-depth studies have examined the effects of the surrounding microenvironment on the survival, proliferation, and expression of markers of DPSC post-neuronal differentiation.…”