利用创新的冷冻细胞库系统高通量体外生产体细胞胚胎

Weiming Wang, Lambert Sanders, Alice Perry, Ebonee Duncan
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摘要

ArborGen公司最初是由West Rock (MWV)、International Paper和Rubicon (Fletcher Challenge)在2000年建立的林业研究合作伙伴关系。它现在生产世界上最多的传统和先进的遗传树苗。在其高级遗传产品中,品种在产量和木材品质方面具有最高水平的遗传性状,如高度,直径生长,抗病性等。为了扩大品种规模,采用体外体细胞胚胎发生(somatic embryogenesis, SE)作为大量繁殖体细胞胚胎的手段。经过多年的研发工作,我们认识到扩大规模的一个关键挑战是缺乏稳定和可预测的生物工艺平台。影响稳定性和可预见性的主要原因包括:难以持续供应良好的起始细胞材料;由于长期接触含有激素的维持介质而存在体细胞无性系变异的风险;以及环境波动造成污染或损失的风险。因此,我们的SE研发计划旨在开发一个强大的生物工艺平台来减少这些变化。我们的主要成就之一是细胞库(CB)系统的开发,该系统为SE生产提供了高质量的起始细胞材料。细胞库系统包括研究细胞库(RCB)和主细胞库(MCB)的创建。细胞库创建后,对储存的细胞进行测试和验证,以确定其胚胎生产的资格。目前,我们已经成功地将细胞库技术整合到一些经济上重要的树种和作物的体细胞胚胎生产中,如松树、云杉、甜桉树、甘蔗、鳄梨和香蕉。通过SE&CB平台,体细胞胚和体细胞苗的产量是一致的,可预测的,并且在商业规模上具有成本效益。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High throughput in vitro production of somatic embryoswith innovative cryo cell banking system
ArborGen Inc. was originally created as a partnership for forestry research in 2000 by West Rock (MWV), International Paper and Rubicon (Fletcher Challenge). It now produces the largest number of both conventional and advanced genetic tree seedlings in the world. Among its advanced genetics offerings, varietals have the highest level of genetic traits in yield and timber quality such as height, diameter growth, disease resistance and etc. To scale up varietals, in vitro process named somatic embryogenesis (SE) is used as a mean to mass propagate SE seedlings. Over the years of R&D efforts, we recognized that a key challenge for scale-up is the lack of a stable and predictable bioprocess platform. The primary causes that impact stability and predictability include a challenge to receive consistent supplies of good starting cell materials, the risk of somaclonal variation due to long term exposure tomaintenance media containing hormones and risk of contamination or loss from environmental fluctuations. Accordingly our SE R&D program has been targeted to develop a robust bioprocess platform to reduce these variabilities. One of our major achievement is the development of a cell banking (CB) system, which provides high quality starting cell materials for SE production. The cell banking system includes the creations of research cell banks (RCB) and master cell banks (MCB). After the cell banks are created, the banked cells are tested and validated for their qualification for embryo production. Currently, we successfully integrated the cell banking technology into somatic embryo production in a number of economically important tree species and crops, such as pine, spruce, sweetgum, sugarcane, avocado and banana. With the SE&CB platform, the somatic embryo & somatic seedling yields are consistent, predictable and are produced cost-efficiently on a commercial scale.
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