This study aimed to propose a simple and practical method for culturing primary rat somatotropic cells in vitro free of pericytes contamination. Rat adenohypophyses were randomly divided into two groups. An improved method was used in group A (digesting adenohypophysis with 0.25% trypsin-EDTA, followed by removing pericytes by double filtration and using serum-free medium for culturing somatotropic cells). The traditional method was used in group B (digesting adenohypophysis with 0.35% collagenase, using serum medium for culturing somatotropic cells, and removing pericytes by changing the culture dish). The numbers and viability of somatotropic cells were higher in group A than in group B after 6 days. GH secretion of somatotropic cells was also higher in group A than in group B. Besides, the pericytes grew rapidly only in group B after 3 days. α-SMA, type I collagen, and type III collagen had weaker expression in group A. Also, the viability of pericytes decreased in group A. The improved method could solve the problem of pericytes contamination, and the culture of primary rat somatotropic cells in vitro was successful. This method can be used for other primary cultures with pericytes contamination.

中文翻译：

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培养来自大鼠腺垂体的促生长细胞的改良方法。

这项研究旨在提出一种简单实用的方法，在体外培养无周细胞污染的原代大鼠生长细胞。大鼠腺垂体被随机分为两组。A组使用了一种改进的方法（用0.25％胰蛋白酶-EDTA消化腺垂体，然后通过双重过滤除去周细胞，并使用无血清培养基培养促生长细胞）。B组采用传统方法（用0.35％胶原酶消化腺垂体，使用血清培养基培养促生长细胞，并通过更换培养皿除去周细胞）。6天后，A组的促生长细胞数量和存活率高于B组。A组的促生长细胞GH分泌也高于B组。此外，周细胞仅在3天后才在B组中迅速生长。α-SMA，I型胶原和III型胶原在A组中的表达较弱。此外，A组中周细胞的活力降低。改进的方法可以解决周细胞污染的问题，并且可以培养大鼠原代生长细胞体外成功。该方法可用于其他具有周细胞污染的原代培养。