Establishment and Characterization of Cell Lines from Primary Culture of Hemangioblastoma Stromal Cells
Establishment and Characterization of Cell Lines from Primary Culture of Hemangioblastoma Stromal Cells
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血管母细胞瘤基质细胞原代培养细胞系的建立和表征
DOI：
10.4103/0028-3886.280643
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发表时间：
2020-03
期刊：
影响因子：
2.7
通讯作者：
Chao You
中科院分区：
文献类型：
--
作者：
Wenke Liu;Si Zhang;Xuhui Hui;Chao You
背景：血管母细胞瘤（hemangioblastomas，HBs）的细胞系还很缺乏。目的：本研究旨在探索一种稳定的HB基质细胞原代细胞系的建立方法，并研究其形态学和分子生物学特性。患者与方法：收集13例患者的乙肝表面抗原标本，建立原代基质细胞系。通过形态学观察、免疫细胞化学染色、NeuN、CD 45、Oligo 2、CD 133、CD 34、GFAP、CD 31、Brachyury、CD 133、CD 34、CD 133、CD 34、CD 45等方法对培养的细胞进行鉴定。结果：11例培养成功，培养成功率为84.6%。细胞传代10代，倍增时间为77.2 ± 5.89h。光镜下可见细胞生长旺盛，呈多边形或三角形，异质性明显。免疫细胞化学染色结果显示，所有培养细胞均表达GFAP、CD 31、NeuN、CD 45和Oligo 2，而Brachyury、CD 133和CD 34均表达。透射电镜证实培养的细胞为基质细胞，具有典型的脂滴。晚期常出现溶酶体自噬现象，无凋亡细胞。结论：选择合适的肿瘤标本、短时间的血管离断、理想的消化时间和营养培养基是建立HB基质细胞原代细胞系的关键。来自von Hippel-Lindau病相关的HB和散发性HB的基质细胞可能起源于胚胎学停滞的成血管细胞。
Context: A well-established cell line of hemangioblastomas (HBs) is still lacking. Aim: This study aims to explore a stable way to establish primary cell lines of HB stromal cells and investigate the morphological and molecular features of these cells. Patients and Methods: Specimens of HBs from 13 patients were collected for establishment of primary cell lines of stromal cells. The details on cell culture were described, and the characterizations of cultured cells were conducted by morphological observation, immunocytochemical staining of inhibin-α, brachyury, CD133, CD34, GFAP, CD31, NeuN, CD45, Oligo2, and transmission electron microscopy. Results: Eleven cases were successfully cultured with a success rate of 84.6%. The cultured cells survived for 10 generations with an estimated doubling time of 77.2 ± 5.89 h. Light microscopy revealed that these cells showed vigorous growth status and presented as polygons or trigons with significant heterogeneity. The immunocytochemical staining showed that inhibin-α, brachyury, CD133, and CD34 were expressed in all the cultured cells, whereas the expression of GFAP, CD31, NeuN, CD45, and Oligo2 was all negative. Transmission electron microscopy confirmed that the cultured cells were stromal cells with typical lipid droplets. The phenomenon of lysosomal autophagy was commonly observed without apoptotic cells in late stage. Conclusion: Appropriate selection of tumor specimens, short duration of devascularization, ideal digestion time, and nutritious medium are critical points for establishment of primary cell line of HB stromal cells. Stromal cells from both von Hippel–Lindau disease-related HBs and sporadic HBs might originate from embryologically arrested hemangioblasts.