Abstract
The neural tube consists of neural progenitors (NPs) that acquire different characteristics during gestation due to patterning factors. However, the influence of such patterning factors on human pluripotent stem cells (hPSCs) during in vitro neural differentiation is often unclear. This study compared neural induction protocols involving in vitro patterning with single SMAD inhibition (SSI), retinoic acid (RA) administration and dual SMAD inhibition (DSI). While the derived NP cells expressed known NP markers, they differed in their NP expression profile and differentiation potential. Cortical neuronal cells generated from 1) SSI NPs exhibited less mature neuronal phenotypes, 2) RA NPs exhibited an increased GABAergic phenotype, and 3) DSI NPs exhibited greater expression of glutamatergic lineage markers. Further, although all NPs generated astrocytes, astrocytes derived from the RA-induced NPs had the highest GFAP expression. Differences between NP populations included differential expression of regional identity markers HOXB4, LBX1, OTX1 and GSX2, which persisted into mature neural cell stages. This study suggests that patterning factors regulate how potential NPs may differentiate into specific neuronal and glial cell types in vitro. This challenges the utility of generic neural induction procedures, while highlighting the importance of carefully selecting specific NP protocols.
| Original language | English |
|---|---|
| Pages (from-to) | 25-34 |
| Number of pages | 10 |
| Journal | Stem Cell Research |
| Volume | 32 |
| Early online date | 23 Aug 2018 |
| DOIs | |
| Publication status | Published - Oct 2018 |
Funding
We thank Matt Clancy for his help with designing the brain-patterning figure in Fig. 5 . We thank Gerbren Jacobs for his excellent technical support. AGN was supported by EU MSCA-ITN CognitionNet ( FP7-PEOPL E -2013-ITN 607508 ). VMH is supported by a ZonMw VIDI research grant ( 91712343 ), an E-Rare Joint Call project ( 9003037601 ), and a European Leukodystrophy Association (ELA) Research Grant ( 2014-012L1 ). Appendix A Supplementary data Supplementary Figure 1: Comparison of NP marker expression between H9-, H1- and human iPSC-derived NPs. Image 5 Supplementary Figure 2: Expression of known regional markers in the three neural induction conditions. Image 4 Supplementary Figure 3: Neuron lineage analysis and quantification. Image 3 Supplementary Figure 4: Glial and regional marker expression in astrocytes generated from the three neural induction conditions. Image 2 Supplementary material 5: Supplementary figure legends and methods. Image 1 Appendix A Supplementary data Supplementary data to this article can be found online at https://doi.org/10.1016/j.scr.2018.08.017 .
| Funders | Funder number |
|---|---|
| European Commission | FP7-PEOPLE-2013-ITN 607508 |
| ZonMw | 91712343, 9003037601 |
| Association Européenne contre les Leucodystrophies | 2014-012L1 |
| Erzincan Üniversitesi |
Keywords
- Astrocytes
- In vitro
- Neural progenitors
- Neurons
- Patterning factors
- Pluripotent stem cells