Wu Bo, Guo Shuzhang, Jiang Tao, Ren Xianjun
Department of Orthopedics, 88th Hospital, Tai'an, Shandong, China.
Neurol Res. 2011 Jul;33(6):593-9. doi: 10.1179/1743132810Y.0000000024.
The pathologic changes of demyelination after spinal cord injury (SCI) significantly impair functional recovery of lesioned spinal cord. At present, transplantation of myelinating cells is regarded as a promising strategy for treating demyelination following SCI. Hence, the In vitro culture and growth, differentiation and proliferation of oligodendrocyte precursor cells (OPCs) were intensively investigated in this study.
In vitro cells from cerebral cortices of neonatal rats were primarily cultured and OPCs were then separated by shaking process and differential adhesion. Following cultured in the conditional medium, growth pattern and differentiation of OPCs were continuously studied by both light microscopy and scanning electron microscopy. Furthermore, maturation of OPCs was detected immunochemically and proliferative ability of OPCs In vitro was also evaluated by methyl thiazolyl tetrazolium (MTT) assay.
The distinct stratification of glial cells usually developed around 9-10 days in the primary culture. The OPCs were found primarily living on the surface of confluent astrocytes and these cells typically displayed the simple appearance of immature cells. Furthermore, the OPCs progressively developed in the conditional medium, and these differentiated cells underwent dramatic changes of morphology and also expressed different specific markers. Moreover, the OPCs also proved by MTT assay to proliferate significantly while cultured In vitro.
Demyelination prevents recovery of neural function following SCI. Demyelination has already become a potential therapeutic target for this insidious and challenging problem. The In vitro culture and biological characteristics of OPCs are fundamental and necessary for further investigation of cell transplantation in vivo. Growth pattern, differentiation and proliferation are very vital for therapeutical effects of OPCs following transplantation after SCI.
脊髓损伤（SCI）后的脱髓鞘病理变化显著损害损伤脊髓的功能恢复。目前，移植髓鞘形成细胞被认为是治疗SCI后脱髓鞘的一种有前景的策略。因此，本研究对少突胶质前体细胞（OPCs）的体外培养、生长、分化和增殖进行了深入研究。
对新生大鼠大脑皮质的体外细胞进行原代培养，然后通过振荡处理和差异黏附分离OPCs。在条件培养基中培养后，通过光学显微镜和扫描电子显微镜持续研究OPCs的生长模式和分化情况。此外，通过免疫化学检测OPCs的成熟情况，并通过甲基噻唑基四氮唑（MTT）法评估OPCs的体外增殖能力。
在原代培养中，通常在9 - 10天左右出现明显的胶质细胞分层。发现OPCs主要存在于汇合的星形胶质细胞表面，这些细胞通常呈现未成熟细胞的简单外观。此外，OPCs在条件培养基中逐渐发育，这些分化的细胞形态发生了显著变化，并表达了不同的特异性标志物。而且，MTT法还证明OPCs在体外培养时显著增殖。
脱髓鞘会阻碍SCI后神经功能的恢复。脱髓鞘已经成为这个隐匿且具有挑战性问题的一个潜在治疗靶点。OPCs的体外培养和生物学特性对于进一步研究体内细胞移植至关重要。生长模式、分化和增殖对于SCI后OPCs移植的治疗效果非常关键。
