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首页> 外文期刊>Laboratory Animal Science >Glucocorticoid-resistant B-lymphoblast cell line derived from the bolivian squirrel monkey (Saimiri boliviensis boliviensis)
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Glucocorticoid-resistant B-lymphoblast cell line derived from the bolivian squirrel monkey (Saimiri boliviensis boliviensis)

机译：来自玻利维亚松鼠猴（Saimiri boliviensis boliviensis）的耐糖皮质激素的B淋巴母细胞细胞系

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The goal of the study reported here was to-develop a continuous cell line from the squirrel monkey that expresses the species-specific phenotype of impaired sensitivity to glucocorticoids. Thirty milliliters of blood from a male Bolivian squirrel monkey (Saimiri boliviensis boliviensis) was fractionated, and the buffy coat was obtained and incubated in the presence of B95-8 cell-conditioned medium, an abundant source of Epstein-Barr virus (EBV), and 2 #mu#g of cyclosporin A/ml. Cell growth was detected within 8 weeks, after which the cells were cloned by use of the limiting dilution method. One clone (4D8) was characterized in detail. The chromosomal count and G-banding pattern confirmed that the cells were of Bolivian squirrel monkey origin. The B-cell origin of these cells was indicated by electron microscopic analysis and was confirmed by expression of CD20. The cells stained strongly for LMP1, a marker of latent EBV infection, and occasionally for the lytic infection marker ZEBRA (BZLF1). The responsiveness of clone 4D8 cells to glucocorticoids was determined by comparing the effects of dexamethasone on cell growth and the induction of a glucocorticoid-inducible mRNA in 4D8 cells with the effects on a human EBV-transformed B-lymphoblast cell line (HL). Dexamethasone inhibited the growth of ilL cells, with IC_(50) of approximately 9 nM, but had no effect on the growth of 4D8 cells. The induction of FK506-binding protein FKBP51 mRNA by dexamethasone was also significantly blunted in 4D8 cells. Thus, we have developed and characterized a squirrel monkey lymphoblastic cell line derived by transformation of B-lymphocytes with EBV; the cell line has diminished growth and transcriptional responses to glucocorticoids.
机译：此处报道的研究目标是从松鼠猴中开发一种连续细胞​​系，该细胞表达对糖皮质激素敏感性降低的物种特异性表型。分离出来自一头玻利维亚松鼠猴（Saimiri boliviensis boliviensis）的30毫升血液，并获得血沉棕黄层，并在B95-8细胞条件培养基（一种丰富的爱泼斯坦-巴尔病毒（EBV）来源）的存在下进行孵育，和2＃μ＃g的环孢菌素A / ml。在8周内检测到细胞生长，然后通过有限稀释法克隆细胞。详细描述了一个克隆（4D8）。染色体计数和G带模式证实该细胞是玻利维亚松鼠猴来源的。这些细胞的B细胞来源通过电子显微镜分析指示，并通过CD20的表达证实。细胞对LMP1（潜在的EBV感染的标志物）进行了强烈染色，偶尔对裂解性感染标志物ZEBRA（BZLF1）进行了染色。通过比较地塞米松对4D8细胞中细胞生长和糖皮质激素诱导型mRNA的诱导作用以及对人EBV转化的B淋巴母细胞系（HL）的影响，确定了克隆4D8细胞对糖皮质激素的反应性。地塞米松抑制ilL细胞的生长，IC_（50）约为9 nM，但对4D8细胞的生长没有影响。地塞米松对FK506结合蛋白FKBP51 mRNA的诱导在4D8细胞中也明显减弱。因此，我们已经开发并鉴定了通过用EBV转化B淋巴细胞衍生的松鼠猴成淋巴细胞细胞系。该细胞系减少了对糖皮质激素的生长和转录反应。

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