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. 2021 Jan-Dec:30:9636897211017829.
doi: 10.1177/09636897211017829.

Differentiation of Human Pluripotent Stem Cells Into Specific Neural Lineages

Chia-Yu Chang  1   2   3 , Hsiao-Chien Ting  1 , Ching-Ann Liu  1   2   3 , Hong-Lin Su  4 , Tzyy-Wen Chiou  5 , Horng-Jyh Harn  1   6 , Shinn-Zong Lin  1   7 , Tsung-Jung Ho  8   9   10
Affiliations

Differentiation of Human Pluripotent Stem Cells Into Specific Neural Lineages

Chia-Yu Chang et al. Cell Transplant. 2021 Jan-Dec.

Abstract

Human pluripotent stem cells (hPSCs) are sources of several somatic cell types for human developmental studies, in vitro disease modeling, and cell transplantation therapy. Improving strategies of derivation of high-purity specific neural and glial lineages from hPSCs is critical for application to the study and therapy of the nervous system. Here, we will focus on the principles behind establishment of neuron and glia differentiation methods according to developmental studies. We will also highlight the limitations and challenges associated with the differentiation of several "difficult" neural lineages and delay in neuronal maturation and functional integration. To overcome these challenges, we will introduce strategies and novel technologies aimed at improving the differentiation of various neural lineages to expand the application potential of hPSCs to the study of the nervous system.

Keywords: central nerve system (CNS); human pluripotent stem cells (hPSCs); induced pluripotent stem cells (iPSCs); neuron and glia differentiation; peripheral nerve system (PNS); specific neural lineages.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Differentiation processes and guidelines of specific neuron and glial cell types from hPSCs. (A) The differentiation steps of specific neurons and glial cells, including NE differentiation, specific neural progenitor patterning, and neuro/glio-genesis. (B) Specific neuronal and glial types can be differentiated from hPSCs under the principles of neural tube development, including the CNS/PNS stem cell differentiation, D-V and A-P determination.
Figure 2.
Figure 2.
Neural patterning principles in neural tube development. Morphogens Wnt, BMP, and Shh are involved in the D-V determination. Wnt and RA regulate the A-P development. Thus, specific neural types are patterned according to their positions in the neural tube.
Figure 3.
Figure 3.
Challenges in obtaining “difficult” neuronal lineages. (A) The lack of in vivo like cell-cell interaction in culture dishes. (B) The major causes that influence the stability of dose dependent patterning protocol. (C) Specialized structures in the CNS.
Figure 4.
Figure 4.
Potential strategies in obtaining “difficult” neuronal lineages. (A) Combination of morphogens to target specific neural lineage. (B) Organoid method for obtaining specialized tissue like structures for specific neural lineages. (C) Sorting and expansion method for high purity interested neural types. (D) Over-expression of specific genes for obtaining interested neural lineages.
Figure 5.
Figure 5.
Application of single cell RNA sequencing on specific neural lineage differentiation from hPSCs. After differentiation of hPSCs into regional specific neurons, single cell RNA sequencing can be applied to cluster the neurons according to their transcriptome. Thus, after transcriptional properties comparison, novel differentiation protocol and surface markers can be identified.
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