Repeated human deciduous tooth-derived dental pulp cell reprogramming factor transfection yields multipotent intermediate cells with enhanced iPS cell formation capability
Repeated human deciduous tooth-derived dental pulp cell reprogramming factor transfection yields multipotent intermediate cells with enhanced iPS cell formation capability
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重复转染人乳牙源性牙髓细胞重编程因子可产生具有增强 iPS 细胞形成能力的多能中间细胞
DOI：
10.1038/s41598-018-37291-2
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发表时间：
2019
影响因子：
4.6
通讯作者：
Sato Masahiro
中科院分区：
文献类型：
--
作者：
Soda Miki;Saitoh Issei;Murakami Tomoya;Inada Emi;Iwase Yoko;Noguchi Hirofumi;Shibasaki Shinji;Kurosawa Mie;Sawami Tadashi;Terunuma Miho;Kubota Naoko;Terao Yutaka;Ohshima Hayato;Hayasaki Haruaki;Sato Masahiro
人体组织特异性干细胞(HTSCs)在体内和体外均可在适当的条件下分化为多个谱系。通过用重编程因子转染终末分化细胞，我们以前从胰腺和肝细胞中产生了诱导的TSCs，这些细胞表现出比IPSCs更多的特性，例如异种移植后非常低的肿瘤形成。我们假设，在IPSC过渡之前的状态下部分重编程的hTSCs可以通过瞬时重编程因子的过度表达从任何末端分化的细胞类型中分离出来。人乳牙髓细胞(HDDPC)和人皮肤成纤维细胞的细胞化学染色显示，尽管在一小部分克隆中，但在三周后，作为干细胞标志物的碱性磷酸酶(ALP)活性增加。重复转染组(≤-3)可产生更高效率的IPSC，与亲本完整的HDDPC相比，HDDPC在两周后表现出更大的多潜能。这些结果表明IPSC技术可用于从HDDPC和成纤维细胞中分离TSCs。通常，ESCs/IPSCs在分化过程中会逐步丧失多潜能表型。我们目前的发现表明，当重复将重编程因子引入分化的细胞(如HDDPC和成纤维细胞)时，也可以发生相反的现象。
Human tissue-specific stem cells (hTSCs), found throughout the body, can differentiate into several lineages under appropriate conditionsin vitroandin vivo. By transfecting terminally differentiated cells with reprogramming factors, we previously produced induced TSCs from the pancreas and hepatocytes that exhibit additional properties than iPSCs, as exemplified by very low tumour formation after xenogenic transplantation. We hypothesised that hTSCs, being partially reprogrammed in a state just prior to iPSC transition, could be isolated from any terminally differentiated cell type through transient reprogramming factor overexpression. Cytochemical staining of human deciduous tooth-derived dental pulp cells (HDDPCs) and human skin-derived fibroblasts following transfection with Yamanaka’s factors demonstrated increased ALP activity, a stem cell marker, three weeks after transfection albeit in a small percentage of clones. Repeated transfections (≤3) led to more efficient iPSC generation, with HDDPCs exhibiting greater multipotentiality at two weeks post-transfection than the parental intact HDDPCs. These results indicated the utility of iPSC technology to isolate TSCs from HDDPCs and fibroblasts. Generally, a step-wise loss of pluripotential phenotypes in ESCs/iPSCs occurs during their differentiation process. Our present findings suggest that the reverse phenomenon can also occur upon repeated introduction of reprogramming factors into differentiated cells such as HDDPCs and fibroblasts.