Establishment of Cultural Method of Primary Rabbit Spinal Cord Microvascular Endothelial Cells
10.13865/j.cnki.cjbmb.2025.04.1076
- VernacularTitle:原代家兔脊髓微血管内皮细胞培养方法的建立
- Author:
Hua-Gen MA
1
;
Ming CHI
;
Zhi-Yi LIN
;
Yuan-Yu TANG
;
Wei-Hong CONG
Author Information
1. 中国中医科学院西苑医院心血管病实验室,北京 100091
- Publication Type:Journal Article
- Keywords:
spinal cord microvascular endothelial cells(SCMECs);
primary culture;
factor Ⅷ-related antigen;
rabbit;
spinal cord injury
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(7):1062-1067
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to isolate and culture primary rabbit spinal cord microvascular endothelial cells in vitro,providing a practical source of test cells for spinal cord injury research.Spinal cord tissue was aseptically extracted from one-month-old rabbits and processed sequentially through mincing,bovine serum albumin density gradient centrifugation,mesh filtration,and type Ⅱ collagenase digestion to ob-tain purified spinal cord microvascular segments.The microvascular segments were homogeneously mixed with an apprapriate volume of M199 complete culture medium and seeded into a culture dish for primary culture.Throughout the culture period,cell growth performance were continuously observed and recor-ded.Additionally,immunocytochemical staining was performed to evaluate the expression of factor Ⅷ-re-lated antigen.The results showed that after 24 hours of inoculation,a small amount of endothelial-like cells were observed to emerge from the spinal cord microvascular segments.Within 36～60 hours,the cell colonies gradually expanded and fused.After 72 hours,the cells spread across the base of the dish,forming a"cobblestone-like"monolayer.Immunocytochemical staining showed that more than 99％of the cells showed brown-red cytoplasm and were positive for factor Ⅷ-related antigen.It is these results that suggest this study has successfully established a convenient and stable primary rabbit spinal cord micro-vascular endothelial cells culture method.
