大鼠松果体细胞体外培养及其生物学特性研究

Study on the culture of rat pinealocyte in vitro and it′s biological characteristics

目的 :了解体外培养条件下松果体细胞的形态、生长、增殖及其生物学特性。方法 :无菌技术手术显微镜下剥离、收集乳鼠松果体 ,胰酶消化分离细胞 ,用含 1 0 %小牛血清的DMEM培养液体外培养 ,倒置显微镜观察 ,四唑盐比色测定细胞活性和增殖 ,Hortega染色及 5 -HT免疫细胞化学法鉴定松果体细胞及其功能状态。结果 :体外培养可获得生长旺盛的松果体细胞 ,细胞的生长倍增时间为第 9d ,第 1 0d达高峰。Hortega染色后松果体细胞呈黑色 ,5 -HT免疫阳性细胞占培养细胞的决大部分。在生存期内 ,阳性细胞数量恒定 ,不因培养时间的延长而改变。结论 :体外培养可获得有增殖潜力的松果体细胞 ,细胞在体外功能活跃 。

Objective: To understand the growth, morphology, proliferation of the rat pinealocytes and it′s biological characteristics in vitro. Methods : The pineal body of neonatal rats was removed by microsurgical technique. The pinealocytes were then isolated by digesting pineals with trypsin. Cells were cultured in DMEM culture medium added with 10% TBS. The vitality of cells was measured by sensitive MTT method and their morphologic features were observed by the inverted phase contract microscope. The special Hortega stain and 5-HT immunocytochemical assay(ABC) were used to detect pinealocytes. Results: Pinealocytes cultured in vitro grew vigorously. The time of cellular group duplication was the ninth day and reached the peak at the tenth day. In the Hortega stained specimen cells were stained black, immunocytochemical staining revealed that the majority of the pinealocytes cultured remained 5-HT positive. During the living period, the number of pinealocytes kept stable. Conclusions: The pinealocytes grow with vitality and keep it′s biological properties in vitro which would not change during the survival period of pinealocytes cultured.