Research progress and application prospect of induced pluripotent stem cells

doi:10.3969/j.issn.2095-4344.2014.01.025

Abstract

Abstract:

BACKGROUND: Induced pluripotent stem cells research has become a hot topic nowadays, because it can avoid the immune rejection and ethical issues in stem cell research and have characteristics is similar to the embryonic stem cells.
OBJECTIVE: To review prospect and progress of induced pluripotent stem cells, and to find out some problems about the induced pluripotent stem cells in inducing method, efficiency, safety and application which need to be solved.
METHODS: Kjmed database was searched by the first author using key words of “induced pluripotent stem cells, efficiency, disease model, clinical application” in English to retrieve relevant articles published from January 2003 to June 2013. Literatures addressing induced pluripotent stem cells were included, and the repetitive researches were excluded.
RESULTS AND CONCLUSION: A total of 112 documents were retieved, and finally 32 articles were retained after deleting unrelated and repetitive ones. Induced pluripotent stem cells have very high application value and prospects in the somatic cell transplantation, tissue engineering, drug screening and establishment in vitro of disease models. But this technology is still in the stage of research and exploration, and there are many technical problems that need to be solved.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, embryonic stem cells, efficiency, transcription factors

CLC Number:

R318

Liu Fei-fei, Hou Zong-liu. Research progress and application prospect of induced pluripotent stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(1): 149-154.

Figures/Tables 1

2.1 纳入文献的基本情况共纳入32篇文献，其中23篇涉及有关诱导性多潜能干细胞诱导方法、筛选鉴定、诱导效率和安全性的研究，4篇涉及诱导性多潜能干细胞在建立疾病模型方面的研究，5篇涉及诱导性多潜能干细胞应用于疾病治疗的研究。 2.2 结果 2.2.1 诱导性多潜能干细胞的建立诱导性多潜能干细胞的诱导方法及诱导效率：通过导入特定的转录因子可将成体细胞重编程为诱导性多能干细胞，最初研究者们应用反转录病毒为载体将转录因子导入成体细胞，但由于此种方法可能使外源基因插入细胞的基因组中引起插入突变，故存在一定的安全隐患；另外，转染效率低也是诱导性多潜能干细胞技术的一个主要障碍，于是研究者们开始探寻一些可以避免基因插入突变并且能够提高诱导效率的新方法。不同诱导方法及诱导剂的探索：Stadtfeld等[2]发现，利用小鼠成纤维细胞，只需含有一个拷贝的Oct4转基因成分，就能通过腺病毒载体将Sox2、c-Myc和Klf4这3个基因转入小鼠成纤维细胞而成功诱导出诱导性多潜能干细胞。Okita等[3]研究者则使用质粒来转染细胞。他们使用2个质粒，一个质粒携带Oct4、Klf4和Sox2基因，而另一个质粒携带c-Myc基因，重复转染小鼠胚胎成纤维细胞7 d后成功诱导出诱导性多潜能干细胞。但和使用腺病毒载体一样，质粒载体的效率相比反转录病毒载体来说更低。并且使用质粒载体也可能出现外源基因随机整合入细胞基因组的情况。随着诱导性多潜能干细胞重编程方法的不断探索，研究者们发现了另一种避免细胞基因组插入突变的办法就是使用小分子物质来替代细胞重编程过程中使用的外源基因，这些小分子物质还能提高转染效率，2009年10月，美国Scripps研究所丁盛博士等发现，在培养液中加入一些小分子化合物如SB43142和PD0325901时，可以显著提高重编程的效率[4]。Huang等[5]在研究中发现只转染2个因子Oct4和Sox2，再加上丙戊酸(valproic acid， VPA)，就能将人类成纤维细胞重编程为诱导性多潜能干细胞。现在人们通过试验发现在人类和小鼠细胞试验中，VPA都可以很大地提高重编程的效率。还有研究者发现在培养过程中添加维生素C可提高iPS重编程效率，它可促进基因表达从而加速体细胞进入重编程状态[6]。由于mRNA、miRNA及siRNA具有不会插入哺乳动物基因组的特点，所以在诱导诱导性多潜能干细胞中有着独特的优势。Warren等[7]将Oct4、Sox2、Klf4、c-Myc这4个关键蛋白质的mRNA导入人皮肤细胞，得到核糖核酸诱导多能干细胞，使细胞重编程的速度提高了1倍，而重编程效率是标准方法的100倍以上，并且没有发生基因组结构改变，核糖核酸诱导多能干细胞比一般的诱导性多潜能干细胞更具备胚胎干细胞的特点。与此同时，有研究用miRNA诱导小鼠和人体细胞生成了诱导性多潜能干细胞，也将重编程效率提高了将近100倍。通过在小鼠胚胎成纤维细胞中表达miR302/367的miRNA簇而获得了类似于诱导性多潜能干细胞的群落，分析其原因，认为可能与其能激活Oct4并抑制HDAC2(组蛋白脱乙酰基酶)的活性有关[8]。 p53是调控细胞程序重排的关键因子，Hong等[9]发现去除c-Myc基因后用siRNA阻断p53基因的路径，可以将皮肤细胞转化为诱导性多潜能干细胞的成功率提高至10%左右，也有研究证明，p53对诱导性多潜能干细胞的转化效率起重要的作用。不同组织来源细胞诱导为诱导性多潜能干细胞的效率比较：不同组织来源的细胞诱导生成诱导性多潜能干细胞的效率也是不相同的[10]，见表1。"

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