CAJ | 학술논문

目的介绍一种新的简便有效的体外培养和纯化雪旺细胞(SCs)方法,以获取足够数量、高纯度的SCs.方法取新生3～5 d的SD乳鼠坐骨神经和臂丛神经,用胶原酶消化后收集细胞进行原代培养,细胞接种浓度为3x105个/mL.在接种72 h内用胰酶分步多次消化进行提纯.结果该方法与1.25 g/L胰酶差速酶消提纯SCs相比,能更有效地将SCs和成纤维细胞分离,经免疫荧光染色显示,SCs的纯度能达到95%以上.结论本方法简便易掌握,可重复性高,能获得足够数量、高纯度的SCs供细胞移植研究.
목적 개소일충신적간편유효적체외배양화순화설왕세포(SCs)방법,이획취족구수량、고순도적SCs.방법 취신생3～5 d적SD유서좌골신경화비총신경,용효원매소화후수집세포진행원대배양,세포접충농도위3x105개/mL.재접충72 h내용이매분보다차소화진행제순.결과해방법여1.25 g/L이매차속매소제순SCs상비,능경유효지장SCs화성섬유세포분리,경면역형광염색현시,SCs적순도능체도95%이상.결론 본방법간편역장악,가중복성고,능획득족구수량、고순도적SCs공세포이식연구.
Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies.