Culture and identification of adult autologous neural stem cells from abandoned brain tissues during surgery in patients with intracranial aneurysm

doi:10.3969/j.issn.2095-4344.0904

Abstract

Abstract:

BACKGROUND: A large number of studies on neural stem cell culture have been carried out in aborted fetus, fetal and adult mice, but there are few studies on adult human neural stem cell culture.
OBJECTIVE: With the improvement of primary cell culture, to culture and identify adult autologous neural stem cells from the abandoned brain tissues around hematoma cavity during surgery in patients with aneurysm rupture, and then to study the cell cryopreservation and recovery so as to establish the adult autologous neural stem cell bank.
METHODS: Over 500 mg abandoned brain tissues were collected during surgery from the surrounding tissues of hematoma cavity in three cases of aneurysm rupture. Adult autologous neural stem cells were cultured in serum-free medium with small tissue piece culture method. Cell growth was investigated by inverted phase contrast microscope. The expression of nestin, the specific antigen of neural stem cells, was confirmed by immunofluorescence method. Culture medium containing 4% fetal bovine serum was used to induce adherent growth of neural stem cells. Subculture was carried out when the cells proliferated to a certain number. Neural stem cells partially cryopreserved were used to establish the adult autologous neural stem cell bank. Recovered cells continued to subculture and induce differentiation. The cell differentiation was induced in the culture medium containing 10% fetal bovine serum. Expression of glial fibrillary acid protein (GFAP; marker for astrocytes), β-tublin (marker for neurons) and SOX10 (marker for oligodendrocytes) was detected by immunofluorescence method.
RESULTS AND CONCLUSION: Nestin-positive neurospheres were successfully obtained from the abandoned brain tissues in the three cases through the improved serum-free medium culture. The cells could be amplified in vitro and handed from generation to generation. When cultured in the medium containing 10% fetal bovine serum, neurospheres were differentiated into β-tublin-positive neurons, SOX10-positive oligodendrocytes and GFAP-positive astrocytes. After subculture, passage 5 cells were still positive for nestin and maintained their characteristics of stemness. After recovery, the cryopreserved cells could grow well, still be positive for Nestin and be passed and amplified continuously. It is proved that neural stem cells can be obtained from abandoned brain tissues in different regions (lower frontal lobe and temporal lobe) around the hematoma cavity in patients with aneurysm rupture. These cells can differentiate into neurons, oligodendrocytes and astrocytes. Autologous neural stem cells transplantation is likely to promote the repair of neural function from laboratory to clinical application.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Tissue Enigneering, Neural Stem Cells, Cells, Cultured, Stem Cells

CLC Number:

R394.2

Wang Jia, Zhao Quan-jun, Hei Bo, Wang Zhao-tao, Sun Yi-kun, Wang Tao, Cui Shao-jie, Wang Pei-xin. Culture and identification of adult autologous neural stem cells from abandoned brain tissues during surgery in patients with intracranial aneurysm[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(21): 3381-3386.

Figures/Tables 1

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