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目的 观察体外培养人嗜铬细胞 (HCC)和微囊化人嗜铬细胞 (ME HCC)的儿茶酚胺(CA)及甲 脑啡肽 (M ENK)的释放功能 ,了解微囊技术对ME HCC形态和活性的影响。方法 取 6例ABO同型健康成人脑死亡的双侧肾上腺进行HCC原代培养 ,对HCC进行HE染色并行免疫细胞化学检查 ,计数酪氨酸羟化酶阳性细胞 ;用 2 %海藻酸钠经微囊技术包裹HCC并进行培养 ,在光镜下观察 ;每 4 8h更换和收集HCC和ME HCC培养液 ,- 2 0℃保存 ;采用放射免疫测定技术检测培养液及烟碱刺激试验后培养液中CA和M ENK水平。结果 (1)ME HCC形态圆整 ,体外培养生长良好 ,与HCC相似 ;(2 )HCC和ME HCC培养液中CA及M ENK水平无显著性差异 (P >0 0 5 ) ;(3)低水平烟碱刺激能提高培养液中HCC和ME -HCC释放CA及M ENK量 ,且二者CA及M ENK增加量无显著性差异 (P>0 0 5 )。结论 本包膜材料和制囊技术对HCC无损伤 ,ME HCC体外培养的活性和释放功能良好 ,适合移植
Objective To observe the release of catecholamine (CA) and me enkephalin (M ENK) from cultured human chromaffin cells (HCC) and microencapsulated human chromaffin cells (ME HCC) Effect of activity. Methods The bilateral adrenal brains from 6 ABO healthy adults with brain death were subjected to HCC primary culture. HCCs were stained with hematoxylin and eosin (HE) for immunocytochemistry and tyrosine hydroxylase positive cells were counted. HCCs were stained with 2% sodium alginate The HCCs were harvested and cultured. The cells were observed under a light microscope. HCC and ME HCC cultures were exchanged and collected every 48 h and stored at -20 ° C. Radioimmunoassay was used to detect CA and M ENK level. Results (1) The morphology of ME HCC was well rounded and grew well in vitro, similar to HCC. (2) There was no significant difference in CA and M ENK between HCC and ME HCC (P> 0.05) Hypobaric nicotine stimulation increased the amount of CA and M ENK released by HCC and ME -HCC in the culture medium, and there was no significant difference in the increase of both CA and M ENK (P> 0.05). Conclusion The envelopment and encapsulation techniques have no damage to HCC, and ME HCC has good activity and release in vitro and is suitable for transplantation