Yukawa Hiroshi, Noguchi Hirofumi, Oishi Koichi, Miyamoto Yoshitaka, Inoue Makoto, Hasegawa Mamoru, Hayashi Shuji, Baba Yoshinobu
Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Higashi-ku, Nagoya, Japan; †FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan.
‡ Baylor All Saints Medical Center and Baylor Research Institute , Dallas, TX , USA.
Cell Med. 2012 May 14;3(1-3):51-61. doi: 10.3727/215517912X639487. eCollection 2012 Jan.
Islet transplantation, including β-cells, has proven to be effective for diabetes in many recent studies; however, this treatment strategy requires sufficient organ donors. One attractive approach for the generation of β-cells is to utilize the expansion and differentiation of cells from pancreatic stem cells (PSCs), which are closely associated to the β-cells lineage. In this study, we investigated whether important transcription factors (Pdx-1, Ngn3, NeuroD, and MafA) in islet cells could be efficiently transduced into mouse PSCs (mPSCs) using Sendai virus (SeV) vectors and found that the transduced cells were differentiated into insulin-producing pancreatic β-cells. The mPSCs transduced with single transcription factors using SeV vectors could not express the insulin-2 mRNA. When combinations of two transcription factors were transduced using the SeV vectors, including combinations of Pdx-1 + NeuroD, Pdx-1 + MafA, and NeuroD + MafA, the expression of insulin-2 mRNA was low but could be detected. When combinations of three or more transcription factors were transduced using SeV vectors, the expression of insulin-2 mRNA could be detected. In particular, the transduction of the combination of PDX-1, NeuroD, and MafA produced the most effective for the expression of insulin-2 mRNA out of all of the different combinations examined. These data suggest that the transduction of transcription factors using SeV vectors facilitates mPSC differentiation into insulin-producing cells and showed the possibility of regenerating β-cells by using transduced PSCs.
包括β细胞在内的胰岛移植在最近的许多研究中已被证明对糖尿病有效;然而,这种治疗策略需要足够的器官供体。一种产生β细胞的有吸引力的方法是利用胰腺干细胞(PSC)来源的细胞的扩增和分化,这些细胞与β细胞谱系密切相关。在本研究中,我们调查了是否可以使用仙台病毒(SeV)载体将胰岛细胞中的重要转录因子(Pdx-1、Ngn3、NeuroD和MafA)有效地转导到小鼠PSC(mPSC)中,并发现转导后的细胞分化为产生胰岛素的胰腺β细胞。使用SeV载体转导单个转录因子的mPSC不能表达胰岛素-2 mRNA。当使用SeV载体转导两个转录因子的组合时,包括Pdx-1 + NeuroD、Pdx-1 + MafA和NeuroD + MafA的组合,胰岛素-2 mRNA的表达较低但可以检测到。当使用SeV载体转导三个或更多转录因子的组合时,可以检测到胰岛素-2 mRNA的表达。特别是,在所有检测的不同组合中,PDX-1、NeuroD和MafA的组合转导对胰岛素-2 mRNA的表达最有效。这些数据表明,使用SeV载体转导转录因子促进了mPSC向产生胰岛素的细胞分化,并显示了通过转导的PSC再生β细胞的可能性。