创伤性脑损伤亚低温治疗期间实施干细胞移植的实验研究

目的 研究在亚低温环境中实施干细胞移植的可行性,为创伤性脑损伤(traumatic brain injuries,TBI)患者接受亚低温治疗期间实施干细胞移植提供实验依据.方法 利用温度敏感型猿猴病毒40大T抗原(tsSV40LT)建立温度敏感型人源性脐带间充质干细胞系(tsUCMSOs),观察其在亚低温(33℃)和正常体温(37℃)下细胞形态、核增殖指数(PIx)及端粒酶活性的变化;建立小鼠TBI亚低温治疗模型,在创伤灶周围半损伤带移植tsUCMSCs,检测其存活率、增殖和凋亡指数,并进行神经功能缺陷评分.结果 tsUCMSCs在33℃培养时,胞体呈长梭形,折光性强,增殖指数和端粒酶活性均较高,但在37℃时胞体扁平,折光性差,细胞老化明显,增值指数和端粒酶活性均明显降低.移植于创伤灶周围半损伤带的tsUCMSCs较对照组(非温度敏感型UCMSCs移植组)细胞存活率明显增高(P<0.05),细胞核增殖抗原高表达,凋亡细胞少见,小鼠神经功能明显好转(P<0.05).结论 tsUCMSCs的建立使TBI亚低温治疗期间实施干细胞移植成为可能,为未来脑创伤的救治提供了新的方法.

Brain stem cell transplantation daring mild hypothermia treatment of patients with traumatic brain injuries

Objective To study the feasibility of stem cell transplantation under mild hypothermia so as to provide a prerequisite for stem cell transplantation in patients with traumatic brain injuries (TBI) during mild hypothermia treatment. Methods After transfecting plasmid containing temperature-sensitive simian virus 40 large T-antigen (tsSV40LT) into temperature-sensitive umbilical cord mes-enchymal stem cells (tsUCMSCs) , the changes of cell morphology, nuclear proliferation index (PIx) and telomerase activity were detested when the tsUCMSCs were cultured at 33℃ and 37℃. After the mouse model with tTBI treated with mild hypothermia was established, tsUCMSCs were transplanted into semi-injury area to detect survival rate, proliferation and apoptosis indices and perform neurological deficit scoring. Results When cultured at 33℃, the tsUCMSCs displayed long spindle-shaped and highly refractive, with higher proliferation index and telomerase activity than those cultured at 37℃. Compared with control group (non-temperature-sensitive UCMSCs transplantation), tsUCMSCs in semi-injury area showed much higher cell survival and proliferating cell nuclear antigen expression ( P < 0.05 ) , with fewer apoptotic cells and better neurological function (P < 0.05). Conclusion The establishment of temperature-sensitive stem cell line enables stem cell transplantation during treatment of TBI with mild hypothermia, as provides us a new direction for treatment of TBI.