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免疫淘选 A2B5 阳性细胞提高了诱导多能干细胞向少突胶质细胞分化的效率。

Immunopanning selection of A2B5-positive cells increased the differentiation efficiency of induced pluripotent stem cells into oligodendrocytes.

机构信息

Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.

出版信息

Neurosci Lett. 2011 Feb 4;489(2):79-83. doi: 10.1016/j.neulet.2010.11.070. Epub 2010 Dec 4.

Abstract

Oligodendrocytes are the myelinating cells of the central nervous system (CNS), and defects in these cells can result in CNS dysfunction. Although oligodendrocyte precursor cell (OPC) transplantation therapy is an effective cure for several disorders, there is no readily available source of these cells. Recent studies have described the generation of induced pluripotent stem cell (iPSC) from somatic cells, leading to speculation that this technique might become a novel therapeutic tool in regenerative medicine. In a previous study, we were able to produce O4 positive (O4(+)) oligodendrocytes from mouse iPSC in vitro. Unfortunately, the efficiency of differentiation achieved was relatively low (2.3%). In the current study, we improved the differentiation efficiency using a mouse monoclonal antibody (A2B5) to select cells of oligodendrocyte lineage. During in vitro differentiation, we purified A2B5-positive (A2B5(+)) cells by immunopanning from a mixed culture of iPSC-derived cells. This procedure increased the differentiation efficiency of O4(+) oligodendrocytes to 43.5%. We also examined the expression of myelin basic protein (MBP), a marker of mature oligodendrocytes. After 21 days of terminal differentiation, 62.3% of iPSC-derived O4(+) oligodendrocytes expressed MBP.

摘要

少突胶质细胞是中枢神经系统 (CNS) 的髓鞘形成细胞,这些细胞的缺陷可导致 CNS 功能障碍。虽然少突胶质前体细胞 (OPC) 移植疗法是治疗几种疾病的有效方法,但目前尚无这些细胞的现成来源。最近的研究描述了从体细胞产生诱导多能干细胞 (iPSC),这引发了人们的猜测,即该技术可能成为再生医学中的一种新型治疗工具。在之前的一项研究中,我们能够在体外从小鼠 iPSC 中产生 O4 阳性 (O4(+)) 少突胶质细胞。不幸的是,达到的分化效率相对较低 (2.3%)。在当前的研究中,我们使用一种小鼠单克隆抗体 (A2B5) 来选择少突胶质细胞谱系的细胞,从而提高了分化效率。在体外分化过程中,我们通过免疫淘选从 iPSC 衍生细胞的混合培养物中纯化 A2B5 阳性 (A2B5(+)) 细胞。这一过程将 O4(+) 少突胶质细胞的分化效率提高到了 43.5%。我们还检查了成熟少突胶质细胞的标志物髓鞘碱性蛋白 (MBP) 的表达。在 21 天的终末分化后,62.3% 的 iPSC 衍生的 O4(+) 少突胶质细胞表达 MBP。

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