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通过核转移技术生成山羊:一项商业运营的回顾性分析(1998-2010 年)。

Generation of goats by nuclear transfer: a retrospective analysis of a commercial operation (1998-2010).

机构信息

LFB USA, Inc., 175 Crossing Boulevard, Framingham, MA, 01702, USA.

出版信息

Transgenic Res. 2020 Aug;29(4):443-459. doi: 10.1007/s11248-020-00207-w. Epub 2020 Jul 1.

Abstract

At LFB USA, Inc., the ultimate use for transgenic cloned goats is for the production of recombinant human protein therapeutics in their milk. This retrospective analysis of the Somatic Cell Nuclear Transfer (SCNT) program, spanning from 1998 to 2010, examined parameters potentially affecting the outcomes and efficiencies in this commercial operation. Over 37,000 + ova were utilized in the SCNT protocol producing a total of 203 cloned goats. Fifty one (51) clones were produced from non-transfected (transgenic and non-transgenic animal donor) cell lines and 152 clones were produced from transfected cell lines. Comparisons and summaries of (a) transfected versus non-transfected cell lines, (b) relationship of SCNT parameters to offspring produced, (c) skin versus fetal cells, (d) fresh versus cryopreserved cells, (e) parameters from all cell lines used versus those producing SCNT offspring, (f) variation among cell sources, (g) methods of SCNT parturition management and effects on live offspring, and lastly (h) SCNT variation by program are reported. Findings indicate that (a) non-transfected cell lines were more efficient versus transfected cell lines in generating viable cloned offspring on a per reconstructed embryo transferred basis, (b) transfected fetal fibroblasts had improved efficiency versus transfected skin fibroblasts, (c) the percentage of non-transfected cell lines that produced offspring was statistically higher than transfected cell lines, (d) and induction of parturition improved the percentage of viable offspring. In summary, this retrospective analysis on the SCNT process has identified certain parameters for improved efficiency in producing viable cloned goats in a commercial setting.

摘要

在美国 LFB 公司,转基因克隆山羊的最终用途是在其奶中生产重组人蛋白治疗药物。本回顾性分析涵盖了从 1998 年到 2010 年的体细胞核移植(SCNT)计划,检查了可能影响商业运营中结果和效率的参数。在 SCNT 方案中使用了超过 37000 个+卵母细胞,共产生了 203 只克隆山羊。51 只克隆羊来自未转染(转基因和非转基因动物供体)细胞系,152 只克隆羊来自转染细胞系。比较和总结了(a)转染与未转染细胞系,(b)SCNT 参数与后代产生的关系,(c)皮肤与胎儿细胞,(d)新鲜与冷冻细胞,(e)所有细胞系的参数与产生 SCNT 后代的参数,(f)细胞来源的变化,(g)SCNT 分娩管理方法及其对活后代的影响,最后(h)报告了该计划的 SCNT 变化。研究结果表明,(a)与转染细胞系相比,未转染细胞系在每转移一个重构胚胎产生可存活的克隆后代方面效率更高,(b)转染胎儿成纤维细胞的效率高于转染皮肤成纤维细胞,(c)产生后代的非转染细胞系的百分比在统计学上高于转染细胞系,(d)诱导分娩提高了活后代的百分比。总之,本回顾性分析 SCNT 过程确定了某些参数,可提高商业环境中生产可存活克隆山羊的效率。

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