一种用于高效体细胞重编程的微流控芯片方法的研究

Reprogramming of somatic cells is to erase the "memory" of somatic cells and reprogramme them to produce new phenotypes and functions. It helps to master the mechanisms of cullular origin and development and to solve the problems in regenerative medicine, as well as has great application value in many fields such as transplantation therapy, illness research, and drug screening. Existing reprogramming methods have their respective defaults, and the fusion of the denucleated embryonic stem cell and somatic cell based method adopts their advantages, which may greatly improve the reprogramming efficiency, so it has numerous potential applications. However, it is still confronted with some difficulties such as low denucleation, fusion and reprogramming efficiency. In this project, microfluidic technology will be used to set up a new method for high-efficiency production of hybrid cells from denucleated embryonic stem cells and somatic cells. Using microfluidic denucleation, alternative sample introduction, dielectrophoretic force based cell pairing, low-voltage electrofusion, and automatic cell sorting, various aspects such as degree of automation, pairing, denucleation efficiency of stem cell, fusion efficiency, cellular activity, and sorting efficiency may be greatly improved and true high-efficiency fusion may be achieved, and it may become a important research tool for reprogramming somatic cells. Meanwhile, many pluripotent cells can be rapidly achieved, which may solve some problems in regenerative medicine such as lack seed cells, and popularize the stem cell transplantation therapy. Thus, this method has great researching significance.

体细胞重编程是指体细胞的"记忆"被擦除，重新程序化产生新表型和功能的方法。其研究有助于掌握机体细胞的发生发育机制，解决再生医学的多个难题，在移植治疗、疾病研究、药物筛选等领域具有重大应用价值。现有重编程方法各有不足，基于去核胚胎干细胞与体细胞融合的方法吸收了它们的优点，有望大大提升重编程效率，因此极具应用前景。但是，它还面临去核效果差，融合及重编程效率不高等难题。 本课题希望利用微流控技术建立一种高效获得去核胚胎干细胞-体细胞融合子的新方法。通过微流控去核操作、交替进样与介电电泳力细胞配对、低压电融合以及细胞自动分离，大大提高自动化程度、干细胞去核率、细胞配对准确率和融合率、融合细胞活性及分离效率，实现真正意义的高效融合，成为体细胞重编程的一种重要研究手段。同时，大量多能细胞的快速获得，有望解决再生医学中种子细胞不足等难题，普及干细胞移植治疗。因此，该方法具有重要研究意义。

体细胞重编程是指体细胞的“记忆”被擦除，重新程序化产生新表型和功能的方法。其研究有助于掌握机体细胞的发生发育机制，解决再生医学的多个难题，在移植治疗、疾病研究、药物筛选等领域具有重大应用价值。现有重编程方法各有不足，基于去核胚胎干细胞与体细胞融合的方法吸收了它们的优点，有望大大提升重编程效率，因此极具应用前景。但是，它还面临去核效果差，融合及重编程效率不高等难题。.经过四年的研究工作，课题组开展了基于离心及化学作用的干细胞去核方法的研究。同时，研究了细胞内容物的释放、富集、收集和脂质体包载，形成了具有细胞内容物的脂质体载体。通过微流控芯片上的细胞和脂（胞）质体的精确控制、配对、电融合，实现了脂（胞）质体和细胞较高效率的融合。针对融合产生的融合子开展了培养分析，证明其具有干细胞的生物学特性。该研究为建立一种更有效的体细胞重编程方法提供了一种新的思路。