Subarachnoid block has no influence on neural differentiation potential of mononuclear cells in the cord blood

doi:10.3969/j.issn.2095-4344.2014.19.014

Abstract

Abstract:

BACKGROUND: Some scholars believe that anesthesia can improve the quality of cord blood separation and collection, and subarachnoid block is commonly used in cesarean section, but there is no research on the influence of subarachnoid block on differentiation potential of mononuclear cells into neural stem cells and astrocytes.
OBJECTIVE: To study the influence of subarachnoid block on differentiation potential of mononuclear cells in the cord blood of neonates into neural stem cells and astrocytes.
METHODS: Mononuclear cells isolated from 20 neonates delivered spontaneously and 20 neonates born by cesarean delivery undergoing subarachnoid block were cultured in vitro, acting as control group and study group. After delivery, cord blood samples were taken to isolate and culture mononuclear cells in vitro. After 3 days of routine culture, the cells were subject to non-induced culture and induced differentiation into neural stem cells. The cultured cells were identified by the cell makers Nestin and glial fibrillary acidic protein with immunohistochemistry identification.
RESULTS AND CONCLUSION: The Nestin and glial fibrillary acidic protein showed no difference between the two groups after induced differentiation (P > 0.05), indicating subarachnoid block has no impact on the differentiation of mononuclear cells in the cord blood from neonates into neural stem cells and astrocytes.

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

全文链接：

Key words: subarachnoid space, anesthesia, obstetrical, fetal blood, glial fibrillary acidic protein, intermediate filament proteins

CLC Number:

R394.2

Zhang Yu-wen, Zhang Xiao-qing, Zhou Ning, Liu Xing-yuan. Subarachnoid block has no influence on neural differentiation potential of mononuclear cells in the cord blood[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(19): 3036-3041.

Figures/Tables 2

2.1 常规培养下脐血单个核细胞形态学观察和生长特点脐血单个核细胞于贴壁前为类圆形细胞，透光度好。接种后24 h左右开始变扁并贴壁。通过体外单层培养及更换为分化培养液时适当洗涤培养板除去不贴壁的红细胞及活性差的细胞等杂细胞，余下生长的细胞为活性好的脐血单个核细胞(图1A，B)。细胞贴培养板壁均匀生长。 2.2 分化培养下细胞形态学观察和生长特点分化培养后细胞逐渐发生形态改变，可见间充质样细胞、破骨样细胞及神经元样细胞出现。通过体外培养，在一定的诱导分化条件下可以使脐血单个核细胞发生显著形态学改变。细胞在分化培养24 h后即开始出现延长，部分呈成纤维样细胞。后续培养过程中，细胞形态渐呈多样化，部分细胞胞浆不规则突起，呈神经元样改变。而继续常规培养的对照组未见明显胞浆突起样细胞。连续动态观察培养板内细胞生长特点：倒置相差显微镜下观察，刚接种的脐血单个核细胞呈小的圆形或椭圆形，大小较均一。接种24 h后可见三四个变扁、贴壁的细胞；刚贴壁时呈单个、分散细胞，接种后72 h大部分细胞贴壁。细胞贴壁后改为分化培养基培养，培养24 h后部分细胞体积增大，部分出现小的胞浆突起，呈圆形、卵圆形或“逗号”样。分化培养72 h后大多数贴壁细胞长出胞浆突起，并可见细胞分裂，研究组更为明显。随着分化培养时间延长，研究组细胞突起增多，部分突起进一步延长，部分长出次级突起，渐呈神经元样细胞。对照组仅大多数表现为细胞变长以梭形细胞为主，胞浆丰富，细胞核大、居中，核仁明显，呈成纤维样细胞形态，平行生长或漩涡生长者居多，培养板中渐出现粒细胞样、破骨细胞样形态不规则细胞。图1-3为倒置相差显微镜下观察不同培养时间培养板内细胞形态。"

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