Shirahata M, Schofield B, Chin B Y, Guilarte T R
Department of Environmental Health Sciences, Johns Hopkins Medical Institutions, Baltimore, MD 21205.
Brain Res. 1994 Sep 26;658(1-2):60-6. doi: 10.1016/s0006-8993(09)90011-7.
Recently patch clamp techniques and optical fluorometric techniques have been applied to freshly dissociated or cultured carotid body. However, very few studies have shown the effects of the dissociation and/or culture conditions on the health and function of the cells. The purpose of this study was to develop a culture method which support healthy and functioning carotid body cells from adult cats. Carotid bodies were dissociated with 0.1-0.2% collagenase and gentle trituration. The cells were plated on glass wells coated with poly-D-lysin and Matrigel, and cultured in chemically defined medium. Culture was maintained for up to 37 days without overgrowth of fibroblasts. Glomus cells extended their processes within and from clusters. Single glomus cells acquired the shape of neurons. Glomus cells synthesized dopamine and its secretion increased during exposure of the cells to hypoxia. Tyrosine hydroxylase was expressed throughout the culture period. These results indicate that glomus cells cultured under conditions described here are healthy and function in a manner similar to that in vivo.
最近，膜片钳技术和光学荧光技术已应用于新鲜解离或培养的颈动脉体。然而，很少有研究表明解离和/或培养条件对细胞健康和功能的影响。本研究的目的是开发一种培养方法，以支持成年猫的健康且功能正常的颈动脉体细胞。颈动脉体用0.1 - 0.2%的胶原酶解离并轻轻研磨。细胞接种于涂有聚-D-赖氨酸和基质胶的玻璃孔中，并在化学成分明确的培养基中培养。培养持续长达37天，成纤维细胞未过度生长。球细胞在细胞簇内并从细胞簇伸出其突起。单个球细胞呈现神经元形状。球细胞合成多巴胺，并且在细胞暴露于缺氧状态时其分泌增加。酪氨酸羟化酶在整个培养期间均有表达。这些结果表明，在此所述条件下培养的球细胞是健康的，并且其功能方式与体内相似。
