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A Defined and Scalable Peptide-Based Platform for the Generation of Human Pluripotent Stem Cell-Derived Astrocytes
journal contribution
posted on 2020-05-20, 15:38 authored by Sreedevi Raman, Gayathri Srinivasan, Nicholas Brookhouser, Toan Nguyen, Tanner Henson, Daylin Morgan, Joshua Cutts, David A. BrafmanAstrocytes
comprise the most abundant cell type in the central
nervous system (CNS) and play critical roles in maintaining neural
tissue homeostasis. In addition, astrocyte dysfunction and death has
been implicated in numerous neurological disorders such as multiple
sclerosis, Alzheimer’s disease, amyotrophic lateral sclerosis
(ALS), and Parkinson’s disease (PD). As such, there is much
interest in using human pluripotent stem cell (hPSC)-derived astrocytes
for drug screening, disease modeling, and regenerative medicine applications.
However, current protocols for generation of astrocytes from hPSCs
are limited by the use of undefined xenogeneic components and two-dimensional
(2D) culture surfaces, which limits their downstream applications
where large-quantities of cells generated under defined conditions
are required. Here, we report the use of a completely synthetic, peptide-based
substrate that allows for the differentiation of highly pure populations
of astrocytes from several independent hPSC lines, including those
derived from patients with neurodegenerative disease. This substrate,
which we demonstrate is compatible with both conventional 2D culture
formats and scalable microcarrier (MC)-based technologies, leads to
the generation of cells that express high levels of canonical astrocytic
markers as well as display properties characteristic of functionally
mature cells including production of apolipoprotein E (ApoE), responsiveness
to inflammatory stimuli, ability to take up amyloid-β (Aβ),
and appearance of robust calcium transients. Finally, we show that
these astrocytes can be cryopreserved without any loss of functionality.
In the future, we anticipate that these methods will enable the development
of bioprocesses for the production of hPSC-derived astrocytes needed
for biomedical research and clinical applications.
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culture surfacesdisease modelingScalable Peptide-Based PlatformPDhPSC linesregenerative medicine applicationsneurodegenerative diseaseapolipoprotein EMCastrocyte dysfunctionamyloid -βHuman Pluripotent Stem Cell-Derived Astrocytes AstrocyteshPSC-derived astrocytestissue homeostasisdisplay propertiesALSxenogeneic componentsCNSpeptide-based substratecanonical astrocytic markersdrug screeningcell typecalcium transientsscalable microcarrier2 D culture formats
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