人鼻黏膜间充质干细胞生物学特性与自体鼻黏膜间充质干细胞移植治疗脊髓损伤

Biological characteristics and clinical observation of autologous nasal mucosa mesenchymal stem cells in the treatment of spinal cord injury

背景:干细胞应用于脊髓损伤的报道很多,但是这些细胞存在免疫排斥、取材困难、纯化不易等各方面的缺陷,而鼻黏膜间充质干细胞无上述缺点,将其应用于临床脊髓损伤患者疗效分析目前尚无相关报道。目的:观察人鼻黏膜间充质干细胞的生物学特性,同时进行自体鼻黏膜间充质干细胞移植治疗晚期不完全性脊髓损伤的临床疗效观察与评价。方法:①取材、培养鼻中隔偏曲手术患者的鼻黏膜间充质干细胞,扫描及透射电子显微镜观察细胞的超微结构,并在体外对其进行多向诱导分化;②经医学伦理委员会同意后,取材培养脊髓损伤患者的鼻黏膜,通过腰椎穿刺术注射到8例晚期不完全性脊髓损伤患者椎管内,细胞移植1-3次,前后移植间隔时间为5-7 d,每次移植细胞总数约为5×107个。细胞移植治疗前及随访6个月时按照国际截瘫医学会ASIA评分标准进行评分。结果与结论:①鼻黏膜间充质干细胞在光镜下形态以梭形为主,并呈现出放射状排列,细胞免疫荧光表达STRO-1;②流式细胞学检测显示高表达间充质干细胞表面标记物CD73、CD90、CD105,不表达造血干细胞标记物CD34、CD45,且细胞纯度达97%以上;③鼻黏膜间充质干细胞在扫描及透射电子显微镜下可见短粗的微绒毛及不同的细胞生长形态;④鼻黏膜间充质干细胞还具有向骨组织、脂肪组织、干细胞球及神经元分化的潜能;⑤经自体血清培养的鼻黏膜间充质干细胞移植后,除1例患者住院期间无明显神经功能改善外,其余患者均有不同程度的神经功能恢复且无任何不良副反应;⑥结果表明,鼻黏膜间充质干细胞是组织工程细胞修复中一种新型的种子细胞来源;自体鼻黏膜间充质干细胞移植治疗脊髓损伤患者能达到一定的治疗效果,具有临床应用价值。

BACKGROUND:Stem cells have been widely used in the treatment of spinal cord injury,but the clinical application islimited by immune rejection,difficulty in cell harvesting and purification.However,human nasal mucosa mesenchymalstem cells(hNM-MSCs)have no such problems,and its clinical application in the treatment of spinal cord injury hasbeen not reported yet.OBJECTIVE:To observe the biological characteristics of autologous hNM-MSCs and its clinical efficacy in thetreatment of advanced incomplete spinal cord injury.METHODS:NM-MSCs were isolated from the human nasal mucosa,cultured and identified in vitro.The ultrastructureof NM-MSCs was observed by transmission electron microscope and scanning electron microscope.Then theNM-MSCs were induced to differentiate into osteocytes,adipocytes,stem cell spheres,or neurons in vitro.Totallyeight patients with incomplete spinal cord injury were enrolled and subjected to hNM-MSCs transplantation via lumbarpuncture for1-3sessions of about5×107cells each,with an interval of5-7days,after the approval of the medicalethics committee.All the patients were followed up for6months.Preoperative and postoperative therapeutic effectevaluations were performed on the basis of American Spinal Injury Association(ASIA)scores.RESULTS AND CONCLUSION:Under light microscope,the NM-MSCs were mainly spindle-shaped,positive forSTRO-1and exhibited a radial arrangement.NM-MSCs highly expressed CD73,CD90and CD105,but did notexpress CD34and CD45,with the purity of over97%.And lots of podgy microvilli were seen on the surface ofNM-MSCs under the scanning electron microscope,and two kinds of cell morphologies were seen under thetransmission electron microscope.Moreover,hNM-MSCs had the ability to differentiate into osteocytes,adipocytes,stem cell spheres and neurons.During the6-month follow-up,seven patients achieved neurological function recoveryto different extents except for one patient,and no side effects were found.It is concluded that hNM-MSCs canbecome the ideal seed cells for tissue-engineered cell repair.Autologous NM-MSCs transplantation for the treatmentof spinal cord injury can achieve the ideal effect,with the value of clinical application.