Induction of functional platelets from mouse and human fibroblasts by p45NF-E2/Maf
Induction of functional platelets from mouse and human fibroblasts by p45NF-E2/Maf
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DOI：
10.1182/blood-2012-02-413617
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发表时间：
2012-11-01
期刊：
影响因子：
20.3
通讯作者：
Matsubara, Yumiko
中科院分区：
文献类型：
--
作者：
Ono, Yukako;Wang, Yuhuan;Matsubara, Yumiko
从干细胞到巨核细胞（MK）和随后的血小板的决定性因素尚未确定。我们现在报道，核因子红细胞衍生的2 p45单位（p45 NF-E2），Maf G和Maf K的组合可以将小鼠成纤维细胞3 T3细胞和成人皮肤成纤维细胞转化为MK。为了筛选MK诱导因子，在不分化成MK的3 T3细胞和已知分化成MK的3 T3-L1细胞之间比较基因表达。在确定的MK谱系诱导培养基中培养用候选因子转染的3 T3细胞。在所测试的因素中，转染p45 NF-E2/MafG/MafK导致CD 41阳性细胞的频率最高。用这些基因转染的成人真皮成纤维细胞在MK谱系诱导培养基中培养。培养的细胞具有巨核细胞特征，包括表面标志物、倍性和形态学。超过90%的MK大小的细胞表达CD 41，称为诱导MK（iMK）。将这些iMK细胞输注到免疫缺陷小鼠中导致CD 41阳性血小板大小颗粒的时间依赖性出现。将来自iMK输注到血小板减少的免疫缺陷小鼠中的血液样品灌注在胶原蛋白包被的芯片上，并且将人CD 41阳性血小板掺入芯片上的血栓中，证明其功能性。这些发现表明p45 NF-E2、Maf G和Maf K的组合是巨核细胞生成和血小板生成的关键决定因素。（血。2012;120（18）：3812-3821）
Determinant factors leading from stem cells to megakaryocytes (MKs) and subsequently platelets have yet to be identified. We now report that a combination of nuclear factor erythroid-derived 2 p45 unit (p45NF-E2), Maf G, and Maf K can convert mouse fibroblast 3T3 cells and adult human dermal fibroblasts into MKs. To screen MK-inducing factors, gene expressions were compared between 3T3 cells that do not differentiate into MKs and 3T3-L1 cells known to differentiate into MKs. 3T3 cells transfected with candidate factors were cultured in a defined MK lineage induction medium. Among the tested factors, transfection with p45NF-E2/MafG/MafK lead to the highest frequency of CD41-positive cells. Adult human dermal fibroblasts transfected with these genes were cultured in MK lineage induction medium. Cultured cells had megakaryocytic features, including surface markers, ploidy, and morphology. More than 90% of MK-sized cells expressed CD41, designated induced MK (iMK). Infusion of these iMK cells into immunodeficient mice led to a time-dependent appearance of CD41-positive, platelet-sized particles. Blood samples from iMK-infused into thrombocytopenic immunodeficient mice were perfused on a collagen-coated chip, and human CD41-positive platelets were incorporated into thrombi on the chip, demonstrating their functionality. These findings demonstrate that a combination of p45NF-E2, Maf G, and Maf K is a key determinant of both megakaryopoiesis and thrombopoiesis. (Blood. 2012;120(18):3812-3821)