BMP inhibition in the presence of LIF differentiates murine embryonic stem cells to early neural stem cells

Title:

BMP inhibition in the presence of LIF differentiates murine embryonic stem cells to early neural stem cells

Creator:

Pisal, R. V. and Mokrý, J.

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:9d8b094f-19a9-4d89-b403-bca901982e55
uuid:9d8b094f-19a9-4d89-b403-bca901982e55

Subject:

early neural stem cells, mouse, neurogenesis, embryonic stem cells, and guided in vitro differentiation

Type:

model:article and TEXT

Format:

bez média and svazek

Description:

Early mouse neural stem cells (NSCs) first appear in embryonic day E5.5 and express pluripotency markers Oct4, Sox2, Nanog and early neural marker Sox1. Early NSCs are a good model for understanding the role of various pathways that control initial neural commitment. However, a protocol for differentiation of mouse embryonic stem cells (ESCs) into early NSCs by adherent monolayer culture has not yet been established. Hence, in this study, we identified the combination of growth factors and small molecules that differentiated mouse ESCs into early NSCs and supported their proliferation. Leukaemia inhibitory factor (LIF) was the first factor to be tested and it was found that ESCs can differentiate into early neurogenic lineage in the presence of LIF. However, we found that the induction is weaker in the presence of LIF as compared to cells differentiated in its absence. GSK-3 inhibitor, along with BMP and TGF-β pathway inhibitor (dual SMAD inhibition), are commonly used to sequentially direct ESCs towards NSCs. However, when we used this combination, mouse ESCs failed to differentiate into early NSCs. We observed that by adding Wnt inhibitor to the combination of GSK-3 inhibitor, BMP inhibitor, TGF-β inhibitor and LIF, it was possible to differentiate ESCs into early NSCs. qRT-PCR analysis of early NSCs illustrated that they expressed key pluripotency genes Oct4 and Nanog, albeit at levels lower than non-differentiated ESCs, along with early neural markers Sox1 and Pax6.

Language:

English

Rights:

http://creativecommons.org/publicdomain/mark/1.0/
policy:public

Coverage:

155-160

Source:

Folia biologica | 2020 Volume:66 | Number:5-6

Harvested from:

CDK

Metadata only:

false