Establishment and characterization of primary astrocyte culture from adult mouse brain

Sun, Xiaoyan; Hu, Xiangyou; Wang, Dan; Yuan, Yihang; Qin, Shangyao; Tan, Zijian; Gu, Yakun; Huang, Xu; He, Cheng; Su, Zhida

doi:10.1016/j.brainresbull.2017.05.002

Cited by 30 publications

(28 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our marker expression data aligns with some aspects of previous studies utilizing primary retinal astrocytes. Other studies report various combinations of GFAP, vimentin, nestin and S100-beta expression in primary retinal and brain astrocytes maintained in serum-containing media (Mi and Barres, 1999;Scheef et al, 2005;Lukas and Wang, 2012;Zamanian et al, 2012;Zhang et al, 2016;Sun et al, 2017). While our detection of vimentin and nestin correlate with these studies, the detection of GFAP does not.…”

Section: Discussionsupporting

confidence: 65%

Phenotypes of primary retinal macroglia: Implications for purification and culture conditions

Backstrom

Sheng

Fischer

et al. 2019

Experimental Eye Research

View full text Add to dashboard Cite

Many neurodegenerations, including those of the visual system, have complex etiologies that include roles for both neurons and glia. In the retina there is evidence that retinal astrocytes play an important role in neurodegeneration. There are several approaches for isolating and growing primary retinal astrocytes, however, they often lead to different results. In this study, we examined the influence of culture conditions on phenotypic maturation of primary, purified retinal glia. We compared retinal astrocytes and Müller glia purified by immunomagnetic separation, as differentiation between these astrocyte subtypes is critical and immuno-based methods are the standard practice of purification. We found that while time in culture impacts the health and phenotype of both astrocytes and Müller glia, the phenotypic maturation of retinal astrocytes was most impacted by serum factors. These factors appeared to actively regulate intermediate filament phenotypes in a manner consistent with the induction of astrocyte-mesenchymal transition (AMT). This propensity for retinal astrocytes to shift along an AMT continuum should be considered when interpreting resulting data. Our goal is that this study will help standardize the field so that studies are replicable, comparable, and as accurate as possible for subsequent interpretation of findings.

show abstract

Section: Discussionsupporting

confidence: 65%

Phenotypes of primary retinal macroglia: Implications for purification and culture conditions

Backstrom

Sheng

Fischer

et al. 2019

Experimental Eye Research

View full text Add to dashboard Cite

show abstract

“…To further demonstrate the molecular heterogeneity of region-restrict astrocytes, we isolated and cultured postnatal astrocytes from the cerebral cortex, cerebellum, and spinal cord. Using a protocol we successfully established recently (Sun et al., 2017), correspondingly, adult astrocytes were also prepared from the three different regions (Figure S1A). Then, the expression level of astrocyte characteristic genes, including Gfap , Aldoc , Glt1 , Glast , Aldh1l1 , CD44 , and Vimentin , was detected in these postnatal or adult astrocytes by real-time qPCR.…”

Section: Resultsmentioning

confidence: 99%

“…Astrocytes can be converted to neuron-like cells by forced expression of transcription factors in vitro (Berninger et al., 2007, Sun et al., 2017). It was reported that a single transcription factor, such as NGN2 or ASCL1, is sufficient to reprogram astrocytes into neurons (Berninger et al., 2007).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Region-Restrict Astrocytes Exhibit Heterogeneous Susceptibility to Neuronal Reprogramming

Qin

Huang

et al. 2019

Stem Cell Reports

Self Cite

View full text Add to dashboard Cite

SummaryThe adult CNS has poor ability to replace degenerated neurons following injury or disease. Recently, direct reprogramming of astrocytes into induced neurons has been proposed as an innovative strategy toward CNS repair. As a cell population that shows high diversity on physiological properties and functions depending on their spatiotemporal distribution, however, whether the astrocyte heterogeneity affect neuronal reprogramming is not clear. Here, we show that astrocytes derived from cortex, cerebellum, and spinal cord exhibit biological heterogeneity and possess distinct susceptibility to transcription factor-induced neuronal reprogramming. The heterogeneous expression level of NOTCH1 signaling in the different CNS regions-derived astrocytes is shown to be responsible for the neuronal reprogramming diversity. Taken together, our findings demonstrate that region-restricted astrocytes reveal different intrinsic limitation of the response to neuronal reprogramming.

show abstract

“…Salsa6f is a genetically encoded ratiometric Ca 2+ indicator derived from the fusion of GCaMP6f and td-Tomato, as described in Dong et al (2017). Adult mouse primary astrocytes were cultured according to the protocol outlined by Sun et al (2017). Cells were dissociated from the cortex, striatum, and hippocampus.…”

Section: Ca 2+ Fluorophores and Tissue Sourcesmentioning

confidence: 99%

Calcium Dynamics in Astrocytes During Cell Injury

Wakida

Gomez-Godinez

et al. 2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

The changes in intracellular calcium concentration ([Ca 2+ ]) following laser-induced cell injury in nearby cells were studied in primary mouse astrocytes selectively expressing the Ca 2+ sensitive GFAP-Cre Salsa6f fluorescent tandem protein, in an Ast1 astrocyte cell line, and in primary mouse astrocytes loaded with Fluo4. Astrocytes in these three systems exhibit distinct changes in [Ca 2+ ] following induced death of nearby cells. Changes in [Ca 2+ ] appear to result from release of Ca 2+ from intracellular organelles, as opposed to influx from the external medium. Salsa6f expressing astrocytes displayed dynamic Ca 2+ changes throughout the phagocytic response, including lamellae protrusion, cytosolic signaling during vesicle formation, vesicle maturation, and vesicle tract formation. Our results demonstrate local changes in [Ca 2+ ] are involved in the process of phagocytosis in astrocytes responding to cell corpses and/or debris.

show abstract

Establishment and characterization of primary astrocyte culture from adult mouse brain

Cited by 30 publications

References 44 publications

Phenotypes of primary retinal macroglia: Implications for purification and culture conditions

Phenotypes of primary retinal macroglia: Implications for purification and culture conditions

Region-Restrict Astrocytes Exhibit Heterogeneous Susceptibility to Neuronal Reprogramming

Calcium Dynamics in Astrocytes During Cell Injury

Contact Info

Product

Resources

About