Immortalization of mouse primary astrocytes

Yi，Weihong1; Yang，Dazhi1; Xu，Zhen1; Chen，Zecai1; Xiao，Guozhi2; Qin，Lei1

2023-05-20

10.1016/j.gene.2023.147327

GENE   Impact Factor & Quartile

0378-1119

1879-0038

In cell culture studies, immortalized primary cells have become a useful tool to investigate the molecular and cellular functions of different types of cells. Several immortalization agents, such as human telomerase reverse transcriptase (hTERT) and Simian Virus 40 (SV40) T antigens, are commonly used for primary cell immortalization. Astrocytes, as the most abundant glial cell type in the central nervous system, are promising therapeutical targets for many neuronal disorders, such as Alzheimer's disease and Parkinson's disease. Immortalized primary astrocytes can provide useful information for astrocytes biology, astrocytes-neuron interactions, glial interactions and astrocytes-associated neuronal diseases. In this study, we successfully purified primary astrocytes with immuno-panning method and examined the astrocyte functions after immortalization through both hTERT and SV40 Large-T antigens. As expected, both immortalized astrocytes presented unlimited lifespan and highly expressed multiple astrocyte-specific markers. However, SV40 Large-T antigen, but not hTERT, immortalized astrocytes displayed fast ATP-induced calcium wave in culture. Hence, SV40 Large-T antigen could be a better choice for primary astrocyte immortalization, which closely mimics the cell biology of primary astrocytes in culture. In summary, the purification and immortalization of primary astrocytes presented in this study can be used for studying astrocyte biology under physiological and pathological conditions.

Astrocytes hTERT antigen Immortalized primary cells SV40 Large-T antigen

English

Corresponding

MOLECULAR BIOLOGY & GENETICS

2-s2.0-85149790886

Scopus

1.Department of Orthopedics,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen,No. 89 Taoyuan Road,518000,China
2.Department of Biochemistry,School of Medicine,Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research,Shenzhen Key Laboratory of Cell Microenvironment,Southern University of Science and Technology,Shenzhen,518055,China

Yi，Weihong,Yang，Dazhi,Xu，Zhen,et al. Immortalization of mouse primary astrocytes[J]. GENE,2023,865.

Yi，Weihong,Yang，Dazhi,Xu，Zhen,Chen，Zecai,Xiao，Guozhi,&Qin，Lei.(2023).Immortalization of mouse primary astrocytes.GENE,865.

Yi，Weihong,et al."Immortalization of mouse primary astrocytes".GENE 865(2023).