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从脐带血管中持续获取内皮细胞和平滑肌细胞培养物的方法。

Procedure to consistently obtain endothelial and smooth muscle cell cultures from umbilical cord vessels.

作者信息

Martín de Llano José Javier, Fuertes Graciela, García-Vicent Consuelo, Torró Isabel, Fayos José Luis, Lurbe Empar

机构信息

Laboratory and the Clinic of the Pediatric Cardiovascular Risk Unit, Pediatric Department, Consorcio Hospital General Universitario de Valencia, Universidad de Valencia, Valencia, Spain.

出版信息

Transl Res. 2007 Jan;149(1):1-9. doi: 10.1016/j.trsl.2006.07.010.

Abstract

The prenatal history of an individual can be responsible to some extent for the occurrence of several diseases later in life. Thus, low birth weight has been related to an increased risk of developing hypertension or type 2 diabetes. The molecular and cellular basis of this increased risk could be found in body fluids and cell types that can be obtained just after birth. To get this unique information, a methodology was developed to consistently obtain cultures of 4 cell types, endothelial and smooth muscle cells from both the vein and the arteries present in the umbilical cord of an individual. From 21 umbilical cords processed, 82 of the 84 possible cell cultures were obtained. The cell cultures exhibit the expected cell morphology and cellular characteristics. Thus, endothelial cells express the von Willebrand factor, CD31, as well as bind and internalize acetylated low-density lipoprotein. Vascular smooth muscle cells express the distinctive alpha-actin. Cell cultures can be cryopreserved and grow healthy for several passages. No influence of birth weight of the newborn has been found in the time required to obtain a primary cell culture for any of the 4 cell types. In conclusion, the procedure developed allows one to routinely obtain actively growing vascular cell cultures that could be used to study the molecular and cellular basis of vascular diseases that emerge in adulthood.

摘要

个体的产前史在一定程度上可能导致其日后患上多种疾病。因此,低出生体重与患高血压或2型糖尿病风险的增加有关。这种风险增加的分子和细胞基础可能存在于出生后即可获取的体液和细胞类型中。为了获取这一独特信息,人们开发了一种方法,能够持续获得4种细胞类型的培养物,即来自个体脐带中静脉和动脉的内皮细胞和平滑肌细胞。在处理的21条脐带中,获得了84种可能的细胞培养物中的82种。这些细胞培养物呈现出预期的细胞形态和细胞特征。因此,内皮细胞表达血管性血友病因子、CD31,并且能结合和内化乙酰化低密度脂蛋白。血管平滑肌细胞表达独特的α-肌动蛋白。细胞培养物可以冷冻保存,并能在传代培养中健康生长。对于这4种细胞类型中的任何一种,在获得原代细胞培养物所需的时间方面,未发现新生儿出生体重有任何影响。总之,所开发的程序使人们能够常规获得活跃生长的血管细胞培养物,这些培养物可用于研究成年期出现的血管疾病的分子和细胞基础。

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