【摘要】 目的探讨成人甲状旁腺细胞的体外培养方法及其分泌功能。方法采用胶原酶法消化成人甲状旁腺腺瘤组织,进行原代和传代细胞培养。每天在显微镜下观察原代细胞和传代细胞的形态变化。测量原代培养1、5、10、15及20 d和传代培养1、5、10及15 d时培养液中甲状旁腺激素(parathyroid hormone,PTH)的水平。结果消化后的细胞形态完整,原代培养2 d时大部分细胞贴壁并展开,3 d时细胞全部展开,4~15 d期间的细胞形态变化不大,16~20 d时部分细胞开始出现老化。传代培养1 d时细胞全部贴壁展开,主要呈梭形,与原代细胞相比体积略有增大;2~15 d期间的细胞形态变化不大。原代细胞上清液中PTH浓度从10 d开始明显降低,15 d时的PTH浓度低于10 d(P<0.01)。同时点传代细胞上清液中PTH的浓度均低于原代甲状旁腺细胞,且5 d和1 d、10 d和5 d、15 d和10 d比较,上清液中PTH浓度的差异均无统计学意义(P>0.05)。结论原代培养10 d内的甲状旁腺细胞具有良好的形态结构,分泌功能最强,可作为永久性甲状旁腺功能低下时同种异体细胞移植的细胞来源;传代后的细胞仍具有分泌功能,但远低于原代细胞。更多还原

【Abstract】 Objective To investigate the way to culture human parathyroid cells and to investigate its secretory function. Methods After digested by collagenase, parathyroid cells were isolated to get the original generation cells, then the cells were cultured and passaged, and morphological changes of original generation cells and passage cells were observed on every day. The parathyroid hormone(PTH) level secreted by the original generation cells and passage cells were measured on the 1st, 5th, 10 th, 15 th, and 20 th day(original generation cells only) respectively. Results The cellular morphology was complete after digestion. On the 2nd day, most of the parathyroid cells had adhered and spreaded, on the 3rd day, all cells had spread. There was no very obvious changes on these cells after cultured for 4-15 days. From 16 to 20 days, some parathyroid cells went senescence. On the 1st day, all of the passage cells, which were fusiform and little bigger than those of the original generation cells, had adhered and spreaded. From 2 to 15 days, there was no very obvious changes. The concentration of PTH in original generation cells begin to decreased significantly on the 10 th day(P<0.01). The concentration of PTH in passage cells were all lower than those of original generation cells at the same corresponding time, but there were no significant difference on the PTH level on 5th day and 1st day, 10 th day and 5th day, 15 th day and 10 th day in passage cells(P>0.05). Conclusions Parathyroid cells which were cultured within 10 days possess well morphologic structure and have the strongest secretory function. Although the passage cells still possess secretory function, it is greatly inferior to original generation cells. At last, we consider that original generation cells cultured within 10 days can be regarded as the source of allogeneic cell transplantation.更多还原