施万细胞与背根节神经元体外共培养成髓鞘模型的建立

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。

Establishment of a Schwann cell-dorsal root ganglion neuron cocluture model of myelination in the peripheral nervous system

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.