用死亡患者源iPSCs研究现代疾病

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用死亡患者源iPSCs研究现代疾病  j0 B, _, ]/ v- R& z5 i
时间：2014年12月4日  来源：生物通
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　　最近，科学家们开发出一种新方法，利用储存在血样中的DNA，重新制备了几十年前去世的患者来源的大脑和肠干细胞，来研究严重疾病（例如炎症性肠道疾病）的潜在原因。这项研究结果发表在最近的美国科学杂志《Stem Cells Translational Medicine》。7 D0 N7 e: A/ O' c
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生物通报道：最近，科学家们开发出一种新方法，利用储存在血样中的DNA，重新制备了几十年前去世的患者来源的大脑和肠干细胞，来研究严重疾病（例如炎症性肠道疾病）的潜在原因。
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这项研究结果发表在最近的美国科学杂志《Stem Cells Translational Medicine》，能够为侵袭性运动神经元和肠道相关疾病患者，产生新的治疗方法，这些疾病已被证明对去世患者是致命的，他们在很久之前自愿捐献出其血样。5 K' c+ ]/ E, `4 N, V
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本文第一作者、再生医学研究所理事会David and Janet Polak 基金会干细胞核心实验室主任、助理教授Dhruv Sareen博士指出：“这项研究的潜在影响是巨大的。”
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通过使用一名死亡患者的储备血样，Sareen及其同事发现，他们能够在培养皿中制备称为iPSCs的干细胞——本质上讲是复活去世很久的患者的患病细胞。2 {; H( g; U1 [4 T* Y' C5 z, \

8 M: o5 n, j3 g) T/ i, l6 ]这一方法允许研究人员把去世患者症状、包含在DNA中的遗传信息和实验室干细胞行为之间联系起来。反过来，这又使研究人员能够研究疾病背后的生物学机制，并可能设计新的治疗方法。
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, o6 {3 `6 d! @6 }该技术还允许医生替换制备生存患者iPSC细胞通常所必需的侵入性活检程序。1 w6 ~* T- M" H9 S* K' K) p5 \6 ^2 P4 T

; ?; ~, ^5 N' G) D3 r再生医学研究所理事会主任Clive Svendsen博士说：“这些新发展使我们能够利用数以百万计患者样本，大规模地制备新的干细胞系。其中一些去世患者被诊断患有罕见和重要的疾病。”
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) d6 H/ i$ l! d& SCedars-Sinai的这项研究，是推动个性化和精确医疗发展、使患者对抗多种疾病的前进一步。克罗恩氏病（Crohn’s disease）是在实验室以这种方法研究的第一种疾病。
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本研究第一作者、再生医学研究所理事会F. Widjaja 基金会炎性肠道和免疫生物学研究所科学家Robert Barrett说：“现在，我们的研究人员可以在培养皿中，制备由罕见突变引起的一种特殊类型克罗恩病患者的完整肠道。这一发展将使我们能够更好地了解，这些突变是如何影响肠道的。”
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原文摘要：
& J8 |3 p/ t, R$ x2 o+ a4 qReliable Generation of Induced Pluripotent Stem Cells From Human Lymphoblastoid Cell Lines0 M) b9 G+ e' ~. I/ q2 N5 y
Abstract: Patient-specific induced pluripotent stem cells (iPSCs) hold great promise for many applications, including disease modeling to elucidate mechanisms involved in disease pathogenesis, drug screening, and ultimately regenerative medicine therapies. A frequently used starting source of cells for reprogramming has been dermal fibroblasts isolated from skin biopsies. However, numerous repositories containing lymphoblastoid cell lines (LCLs) generated from a wide array of patients also exist in abundance. To date, this rich bioresource has been severely underused for iPSC generation. We first attempted to create iPSCs from LCLs using two existing methods but were unsuccessful. Here we report a new and more reliable method for LCL reprogramming using episomal plasmids expressing pluripotency factors and p53 shRNA in combination with small molecules. The LCL-derived iPSCs (LCL-iPSCs) exhibited identical characteristics to fibroblast-derived iPSCs (fib-iPSCs), wherein they retained their genotype, exhibited a normal pluripotency profile, and readily differentiated into all three germ-layer cell types. As expected, they also maintained rearrangement of the heavy chain immunoglobulin locus. Importantly, we also show efficient iPSC generation from LCLs of patients with spinal muscular atrophy and inflammatory bowel disease. These LCL-iPSCs retained the disease mutation and could differentiate into neurons, spinal motor neurons, and intestinal organoids, all of which were virtually indistinguishable from differentiated cells derived from fib-iPSCs. This method for reliably deriving iPSCs from patient LCLs paves the way for using invaluable worldwide LCL repositories to generate new human iPSC lines, thus providing an enormous bioresource for disease modeling, drug discovery, and regenerative medicine applications.