Isolation and Identification of Spinal Astrocytes from Tree Shrews

doi:10.3969/j.issn.1674-5817.2019.01.004

Abstract

Abstract: Objective To establish an effective method of isolation, identification and primary culture method for astrocytes derived from the spinal cord of tree shrews in vitro, providing experimental materials for in vitro studies of spinal astrocytes. Methods Neonatal tree shrews were euthanized to collect spinal cord tissue, the spinal cord membrane and blood vessels were removed under a dissecting microscope to reduce the effects of fibroblasts, vascular endothelial cells and red blood cells. Then, the spinal cord tissues were digested with trypsin and DNase I, and cultured. According to the difference in the adherence time of astrocytes, fibroblasts and microglia, the fibroblasts were removed by differential adherence speed. After the cells were confluent, the non-staple gel was removed by shaking with a constant temperature shaker. Neurons were removed by three consecutive purifications. The cultured cells were identified by immunofluorescence staining of glial fibrillary acidic protein (GFAP). Results The astrocytes were isolated by differential adherent culture after trypsin and DNaseI digestion. The isolated cells had typical protoplasmic and fibrous characteristics, the protoplasmic type was polygonal, the protrusions were few and strong, the fibrous protrusions are abundant and spread outward. The cellular immunofluorescence results showed that GFAP expression is positive, and the purity could reach 95%. Conclusion The isolation and purification of spinal cord astrocytes from tree shrews are successfully established.

Key words: Astrocytes, Spinal cord, Isolation and identification, Tree shrews

CLC Number:

Q95-33

WANG Xuan, WANG Wen-guang, LI Na, YUAN Yuan, ZHANG Zhi-cheng, SUN Xiao-mei. Isolation and Identification of Spinal Astrocytes from Tree Shrews[J]. Laboratory Animal and Comparative Medicine, 2019, 39(1): 15-20.

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