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曼氏血吸虫幼体蠕虫细胞的培养

Culture of cells from juvenile worms of Schistosoma mansoni.

作者信息

Hobbs D J, Fryer S E, Duimstra J R, Hedstrom O R, Brodie A E, Collodi P A, Menino J S, Bayne C J, Barnes D W

机构信息

Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.

出版信息

J Parasitol. 1993 Dec;79(6):913-21.

PMID:8277385
Abstract

Tissue disruption methods were developed and serum-free cell culture media formulated for the maintenance in vitro of cells from juvenile worms (day 18 after infection) of Schistosoma mansoni. Cultures maintained viability for up to 6 mo when plated on a feeder layer of irradiated rat liver cells and survived primarily as clusters of small (2.5-4 microns diameter) cells with a high nuclear-to-cytoplasmic ratio and relatively few organelles identified by electron microscopy. Cultures synthesized a protein profile similar to that of intact worms, and the cell clusters maintained a time- and concentration-dependent contractile response to serotonin. Cells synthesizing DNA were detected by precursor incorporation and flow cytometry in cultures initially and also after several weeks in vitro, although the percentage of cells synthesizing DNA decreased with time. Efforts to identify peptide growth factor-responsive tyrosine phosphorylation were negative, and the overall amount of S. mansoni phosphotyrosine-containing proteins identified by western blot with anti-phosphotyrosine monoclonal antibody was much less than that found in a peptide growth factor-responsive mouse cell line.

摘要

开发了组织破坏方法,并配制了无血清细胞培养基,用于体外培养曼氏血吸虫幼虫(感染后第18天)的细胞。当接种到经辐照的大鼠肝细胞饲养层上时,培养物的活力可维持长达6个月,主要以小细胞(直径2.5 - 4微米)簇的形式存活,核质比高,通过电子显微镜鉴定细胞器相对较少。培养物合成的蛋白质谱与完整蠕虫相似,细胞簇对5-羟色胺保持时间和浓度依赖性收缩反应。通过前体掺入和流式细胞术在最初培养的细胞以及体外培养数周后的细胞中检测到了合成DNA的细胞,尽管合成DNA的细胞百分比随时间下降。鉴定肽生长因子反应性酪氨酸磷酸化的努力为阴性,用抗磷酸酪氨酸单克隆抗体通过蛋白质印迹法鉴定的曼氏血吸虫含磷酸酪氨酸蛋白的总量远低于在肽生长因子反应性小鼠细胞系中发现的量。

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