CAJ | 학술논문

目的观察体外培养的微囊化牛嗜铬细胞及其冷冻保存复苏后的细胞活力,探索微囊化牛嗜铬细胞的冻存保存方法,为将来进行大规模细胞移植治疗疼痛奠定临床基础.方法经胶原酶消化成单细胞,微囊包裹,并取部分进行冷冻保存及复温,观察细胞的生长情况;放免法分别检测其培养液中去甲肾上腺素、甲-脑啡呔的含量和在高钾、乙酰胆碱刺激下的分泌量来观察其功能活性.结果微囊化牛嗜铬细胞及其冷冻复苏后的细胞形态结构完整,生长良好;冻存复苏后的微囊保留了儿茶酚胺和甲-脑啡呔的的分泌功能,基础与刺激分泌量大约为冻存前微囊化牛嗜铬细胞分泌量的92%;同时刺激后去甲肾上腺素、甲-脑啡呔水平较其基础值均有明显增加(P＜0.01).结论微囊化牛嗜铬细胞及其冷冻复苏后的细胞具有良好的细胞功能,冷冻保存的微囊化牛嗜铬细胞是有效和成功的.
목적 관찰체외배양적미낭화우기락세포급기냉동보존복소후적세포활력,탐색미낭화우기락세포적동존보존방법,위장래진행대규모세포이식치료동통전정림상기출.방법 경효원매소화성단세포,미낭포과,병취부분진행냉동보존급복온,관찰세포적생장정황;방면법분별검측기배양액중거갑신상선소、갑-뇌배태적함량화재고갑、을선담감자격하적분비량래관찰기공능활성.결과 미낭화우기락세포급기냉동복소후적세포형태결구완정,생장량호;동존복소후적미낭보류료인다분알화갑-뇌배태적적분비공능,기출여자격분비량대약위동존전미낭화우기락세포분비량적92%;동시자격후거갑신상선소、갑-뇌배태수평교기기출치균유명현증가(P＜0.01).결론 미낭화우기락세포급기냉동복소후적세포구유량호적세포공능,냉동보존적미낭화우기락세포시유효화성공적.
Objective To observe the viability of microencapsulated bovine chromaffin cells and their viability after cryopreservation, explore a cryopreservation mean for the microencapsulated chomaffin cells, and build a clinical foundation for the future large-scale clinical cell transplantation for treatment of pain. Methods Bovine adrenal medullary was digested into single chromaffin with collagenase digestion, then microencapsulated it. The cells were cryopreservated and some of them were thawed for in vitro culture to observe the cell growth; Culture medium was collected for the analysis of norepinephrine and methyl-endomorphin concentration by radioimmunoassay. Cell viability was evaluated by the secretion under high potassium and acetylcholine stimulation. Results Microencapsulated bovine chromaffin cells displayed their morphology structural integrity and functioned well after cryopreservation. The cells maintained the ability to secrete catecholamines and methyl-endomorphin. The basic and stimulated secretion volume of microencapsulated bovine chromaffin cells recovered from cryopreservation were 92% of that of fresh chromaffin cells. The norepinephrine and methyl-endomorphin levels were significantly higher after stimulation (P＜0.01). Conclusion Microencapsulated bovine chromaffin cells and thawed cells from cryopreservation show the ability to secret both catecholamines and methyl-endomorphin well, which demonstrated that the microencapsulation and cryopreservation are useful method to preserve bovine chromaffin cells.