CAJ | 학술논문

背景：目前，大量文献报道了诱导多能性干细胞系的建立，但大规模体外诱导分化造血祖细胞的研究还缺乏深入的探讨。 　　目的：建立诱导多能性干细胞体外定向诱导形成造血祖细胞的方法。 　　方法：采用慢病毒感染的方法将含有Oct4、Sox2、Nanog和Lin28全能性基因的慢病毒颗粒转导人皮肤成纤维细胞，获得了诱导多能性干细胞；在诱导分化体系中添加了 Y-27632，克服干细胞扩增中的凋亡现象；运用OP9细胞产生的条件培养液建立诱导多能性干细胞体外定向分化形成造血祖细胞的分化体系。 　　结果与结论：①前3代细胞克隆传代时，诱导多能性干细胞发生凋亡的现象很多，很难大规模扩增培养。培养基中添加阻断 ROCK活化的抑制剂，能够明显抑制胚胎干细胞的凋亡。②诱导多能性干细胞在 OP9细胞条件培养液作用下，经过体外诱导分化，形成CD34+造血祖细胞。
배경：목전，대량문헌보도료유도다능성간세포계적건립，단대규모체외유도분화조혈조세포적연구환결핍심입적탐토。 　　목적：건립유도다능성간세포체외정향유도형성조혈조세포적방법。 　　방법：채용만병독감염적방법장함유Oct4、Sox2、Nanog화Lin28전능성기인적만병독과립전도인피부성섬유세포，획득료유도다능성간세포；재유도분화체계중첨가료 Y-27632，극복간세포확증중적조망현상；운용OP9세포산생적조건배양액건립유도다능성간세포체외정향분화형성조혈조세포적분화체계。 　　결과여결론：①전3대세포극륭전대시，유도다능성간세포발생조망적현상흔다，흔난대규모확증배양。배양기중첨가조단 ROCK활화적억제제，능구명현억제배태간세포적조망。②유도다능성간세포재 OP9세포조건배양액작용하，경과체외유도분화，형성CD34+조혈조세포。
BACKGROUND:There have been a large number of reports on establishing induced pluripotent stem celllines, but studies concerning large-scale in vitro induced differentiation of induced pluripotent stem cels into hematopoietic progenitor cels stil have a lack of in-depth discussion. OBJECTIVE:To develop methods to induce differentiation of induced pluripotent stem cels into hematopoietic progenitor cels in vitro. METHODS: Using the method of infection with lentivirus particles containing four transcriptionfactor genes, which are Oct4, Sox2, Nanog and Lin28, human skin fibroblasts are transduced into induced pluripotent stem cels. In the induced differentiation system, Y-27632, a kind of tyrosine kinase inhibitor-ROCK (p160-Rho-associated coiled-coil kinase), was added, which obviously suppressed apoptosis of cels. Based on conditioned medium with OP9 cels, a differentiation system of inducing induced pluripotent stem cels differentiating into hematopoietic progenitor cels was established. RESULTS AND CONCLUSION:(1) Apoptosis of induced pluripotent stem cels at the first three passages was very obvious, and the cels were difficult in a large-scale expansion. After Y-27632 was added, the apoptosis of embryonic stem cels was obviously inhibited. (2) During embryoid body differentiation, induced pluripotent stem cels cultured in OP9 conditional growth medium differentiated into hematopoietic progenitor celsin vitro that were positive for CD34.