CAJ | 학술논문

目的：探讨施万细胞与背根节神经元髓鞘化共培养的标准化方法，为研究周围神经髓鞘化的形成机制提供稳定的周围神经髓鞘化体外模型。方法：取出生1~3 d新生SD大鼠，培养施万细胞，经纯化鉴定后用于共培养。取孕14~15 d的SD大鼠胚鼠背根神经节，经纯化后用于共培养；将2种分别纯化的细胞进行共培养，加抗坏血酸诱导髓鞘的形成。利用免疫组化染色、扫描电镜和透射电镜，检测髓鞘的形成。结果：纯化后施万细胞纯度可达到98%以上，可用于共培养。背根节神经元贴壁良好，经纯化后几乎无杂细胞可见，可用于共培养。对共培养细胞进行MAG与NF的免疫组化染色，发现有数量可观的髓鞘形成，并且髓鞘是紧密包绕在神经元轴突上。扫描电镜下可观察到施万细胞包绕轴突形成髓鞘，而透射电镜下则可观察到有致密的髓鞘板层结构形成。结论：本实验成功建立稳定可靠的体外成髓鞘模型，可用于轴突的新髓鞘化实验研究。
목적：탐토시만세포여배근절신경원수초화공배양적표준화방법，위연구주위신경수초화적형성궤제제공은정적주위신경수초화체외모형。방법：취출생1~3 d신생SD대서，배양시만세포，경순화감정후용우공배양。취잉14~15 d적SD대서배서배근신경절，경순화후용우공배양；장2충분별순화적세포진행공배양，가항배혈산유도수초적형성。이용면역조화염색、소묘전경화투사전경，검측수초적형성。결과：순화후시만세포순도가체도98%이상，가용우공배양。배근절신경원첩벽량호，경순화후궤호무잡세포가견，가용우공배양。대공배양세포진행MAG여NF적면역조화염색，발현유수량가관적수초형성，병차수초시긴밀포요재신경원축돌상。소묘전경하가관찰도시만세포포요축돌형성수초，이투사전경하칙가관찰도유치밀적수초판층결구형성。결론：본실험성공건립은정가고적체외성수초모형，가용우축돌적신수초화실험연구。
Objective:To study the mechanism of peripheral nerve myeli nation, establish a standardized method of Schwann cell and dorsal root ganglion neuron co-culture system of myelination in vitro were established.Methods:SCs were isolated from the sciatic nerves of newborn rat pups (within postnatal 1~3 d). Dorsal root ganglia (DRG) neurons were isolat-ed from the pups of E14~15d SD rats and through the explant culture of DRG. After purification and qualification, these cells were used for co-culture. Then myelin formation were tested by methods of Immune Fluorescent Staining, scanning electron microscopy and transmission electron microscopy.Results:SCs were arranged neatly and their bodies were uniform after purification.The statistical data of immunofluorescence staining showed that the cell purification could reach up to 98%and they could be used for co-culture; DRG neurons were attached to the cell plate steadily, and no hybrid cells could be observed.The immunofluorescence staining of MAG and NF demonstrated that a significant quantity of myelin had formed, and the myelin surrounded the axons tightly; under the scanning electron microscope the process of myelination were ob-seved while under the transmission electron microscopy showed the typical compact myelin lamellar structure. Conclusions:A stable and reliable model of myelination in vitro has been established and could be used in our subsequent study.