Growth kinetics, plasticity and characterization of hamster embryonic fibroblast cells

D. Mehrabani, N. Booyash, H. Aqababa, Amin Tamadon, S. Zare, M. Dianatpour
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引用次数: 7

Abstract

Abstract Development of different embryonic stem cells (ESCs) requires feeder fibroblast cell culture. Moreover, the establishment of fibroblast cell culture especially for endangered species can provide an excellent resource for biological research and preserve precious genetic materials. This study aimed to characterize and determine the growth kinetic of hamster embryonic fibroblast cells. Hamster fetuses of two albino female hamsters were collected between 8 and 10 days of pregnancy. After removal of the head, liver and gut, the fetuses were cut into 1-mm2 pieces and then cultured. After reaching 80–90% confluence, the cells were subcultured. The cells of passage 8 were subcultured in two 24-well plates (2 × 104 cells/well) for 7 days. Three wells per day were counted and the average cell counts at each time point were plotted against time, and the population doubling time (PDT) was determined. Cell viability after freezing and thawing was evaluated. For karyotyping, the cells of the passage 8 were used. The PDT of the cells at passage 8 was about 34.9 h and the viability was 77.9% in the passage 8. The isolated cells were spindle shaped and plastic adherent. The chromosome number and morphology were normal. The favorable morphology, viability, growth kinetic and karyotyping of hamster embryonic fibroblast cells revealed that these cells even at the eighth passage can safely be used as a feeder layer for ESCs, transgenic purposes and gene banks, and also for biological and pharmacological research.
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仓鼠胚胎成纤维细胞的生长动力学、可塑性和特性
不同胚胎干细胞(ESCs)的发育需要培养成纤维细胞。此外,建立濒危物种成纤维细胞培养体系可以为生物研究提供良好的资源,保存珍贵的遗传物质。本研究旨在表征和确定仓鼠胚胎成纤维细胞的生长动力学。在怀孕8至10天期间收集了两只白化雌性仓鼠的仓鼠胎儿。去头、去肝、去肠后,将胎儿切成1mm2块进行培养。达到80-90%汇合后,细胞传代培养。第8代细胞在2个24孔板(2 × 104个/孔)中传代培养7 d。每天对3口井进行计数,并绘制每个时间点的平均细胞计数与时间的关系图,确定种群倍增时间(PDT)。评估冷冻和解冻后的细胞活力。用第8代的细胞进行核型分析。第8代细胞PDT约为34.9 h,存活率为77.9%。离体细胞呈纺锤形,可塑贴壁。染色体数目和形态正常。小鼠胚胎成纤维细胞的形态、活力、生长动力学和核型分析表明,即使在第8代,这些细胞也可以安全地用作胚胎干细胞、转基因目的和基因库的饲养层,并可用于生物学和药理学研究。
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Italian Journal of Zoology
Italian Journal of Zoology 生物-动物学
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6-12 weeks
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