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无血清器官培养中的人胎儿肾外植体

Human foetal kidney explants in serum-free organ culture.

作者信息

Brière N

出版信息

Anat Embryol (Berl). 1987;176(1):105-14. doi: 10.1007/BF00309758.

Abstract

The purpose of the work was to develop an in vitro model for the study of human kidney development. Human metanephric explants from foetuses 10-18 weeks of gestational age were cultured in serum-free Leibovitz L-15 medium without hormones. Under the current minimal conditions for growth, the system permitted to maintain the renal tissues in culture for periods up to 9 days, although no evident sign of morphological differentiation was observed. However, during the studied period the overall architecture of the explants was preserved as well as the ultrastructural features of cytoplasmic organelles. The incorporation of 3H-thymidine and 3H-leucine indicated that DNA and protein synthesis was maintained or increased. Glycoprotein synthesis evaluated by 3H-glucosamine incorporation and radioautography continued in mesangium as well as in glomerular and tubular basement membranes. Alkaline phosphatase, gamma-glutamyltranspeptidase (brush border) and catalase (peroxisomes) activities remained histochemically active. The proposed organ culture system appears as a reliable and promising model that will provide basic data on the morphology and functional characteristics of the developing kidney. Since it is achieved in a completely controlled environment, it will permit to study the role of growth factors and hormones in proliferation and differentiation of the cell populations during development of the human foetal kidney.

摘要

这项工作的目的是开发一种用于研究人类肾脏发育的体外模型。将孕龄10 - 18周胎儿的人类后肾外植体在无激素的无血清Leibovitz L - 15培养基中培养。在当前的最小生长条件下,该系统可使肾组织在培养中维持长达9天,尽管未观察到明显的形态分化迹象。然而,在研究期间,外植体的整体结构以及细胞质细胞器的超微结构特征得以保留。3H - 胸腺嘧啶核苷和3H - 亮氨酸的掺入表明DNA和蛋白质合成得以维持或增加。通过3H - 葡糖胺掺入和放射自显影评估的糖蛋白合成在系膜以及肾小球和肾小管基底膜中持续进行。碱性磷酸酶、γ - 谷氨酰转肽酶(刷状缘)和过氧化氢酶(过氧化物酶体)活性在组织化学上保持活跃。所提出的器官培养系统似乎是一个可靠且有前景的模型,它将提供有关发育中肾脏的形态和功能特征的基础数据。由于它是在完全受控的环境中实现的,这将有助于研究生长因子和激素在人类胎儿肾脏发育过程中细胞群体增殖和分化中的作用。

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