Functional and phenotypic differences of pure populations of stem cell‐derived astrocytes and neuronal precursor cells. Issue 5 (22nd December 2015)

• Record Type: Journal Article
• Authors: Kleiderman, Susanne; Sá, João V.; Teixeira, Ana P.; Brito, Catarina; Gutbier, Simon; Evje, Lars G.; Hadera, Mussie G.; Glaab, Enrico; Henry, Margit; Sachinidis, Agapios; Alves, Paula M.; Sonnewald, Ursula; Leist, Marcel
• Is Part Of: Glia. Volume 64:Issue 5(2016:May)
• Page Start: 695
• Page End: 715
• Publication Date: 2015-12-22
• Subjects: astrocytes -- neural stem cells -- differentiation -- metabolic flux -- transcriptome; Neuroglia -- Periodicals; Neurology -- Periodicals; 611.0188
• DOI: 10.1002/glia.22954
• Languages: English
• ISSNs: 0894-1491

Abstract

Abstract : Availability of homogeneous astrocyte populations would facilitate research concerning cell plasticity (metabolic and transcriptional adaptations; innate immune responses) and cell cycle reactivation. Current protocols to prepare astrocyte cultures differ in their final content of immature precursor cells, preactivated cells or entirely different cell types. A new method taking care of all these issues would improve research on astrocyte functions. We found here that the exposure of a defined population of pluripotent stem cell‐derived neural stem cells (NSC) to BMP4 results in pure, nonproliferating astrocyte cultures within 24–48 h. These murine astrocytes generated from embryonic stem cells (mAGES) expressed the positive markers GFAP, aquaporin 4 and GLT‐1, supported neuronal function, and acquired innate immune functions such as the response to tumor necrosis factor and interleukin 1. The protocol was applicable to several normal or disease‐prone pluripotent cell lines, and the corresponding mAGES all exited the cell cycle and lost most of their nestin expression, in contrast to astrocytes generated by serum‐addition or obtained as primary cultures. Comparative gene expression analysis of mAGES and NSC allowed quantification of differences between the two cell types and a definition of an improved marker set to define astrocytes. Inclusion of several published data sets in this transcriptome comparison revealed the similarity of mAGES with cortical astrocytes in vivo . Metabolic analysis of homogeneous NSC and astrocyte populations revealed distinct neurochemical features: both cell types synthesized glutamine and citrate, but only mature astrocytes released these metabolites. Thus, the homogeneous cultures allowed an improved definition of NSC and astrocyte features. GLIA 2016;64:695–715 Main points: Rapid generation of a homogeneous population of mature, non‐dividing astrocytes. Comparison of embryonic stem cell‐derived astrocytes (mAGES) and neural stem cells (NSC). Differences of NSC and mAGES concerning function, metabolism, and transcriptome.