Development of feeder-free culture systems for generation of ckit+sca1+ progenitors from mouse iPS cells.
Development of feeder-free culture systems for generation of ckit+sca1+ progenitors from mouse iPS cells.
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DOI：
10.1007/s12015-010-9215-5
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发表时间：
2011-09
影响因子：
4.8
通讯作者：
Roy, Krishnendu
中科院分区：
文献类型：
--
作者：
Lin, Jian;Fernandez, Irina;Roy, Krishnendu
来自诱导多能干细胞(IPS)的患者特异性治疗细胞可能绕过与胚胎干细胞(ES)相关的伦理问题，并避免与同种异体移植相关的潜在免疫反应。对于iPS细胞衍生疗法的最终临床适用性来说，建立无滋养细胞培养以确保iPS细胞高效分化为治疗性前体细胞是至关重要的。还有必要了解iPS细胞来源的祖细胞与ES细胞来源的祖细胞是否不同。在这项研究中，我们比较了三种不同的无饲养层培养对小鼠iPS细胞分化为cKit+SCA1+造血祖细胞(HPC)的效率，并比较了ES细胞和iPS细胞之间分化和功能的差异。我们的结果表明，在含有小鼠骨髓基质细胞分泌因子的悬浮培养中(OP9-DL1条件培养液)，iPS和ES细胞都能有效地分化为HPC。通过向CD11c+树突状细胞(DC)的分化，证实了这些细胞的功能。在混合淋巴细胞反应(MLR)中，ES和iPS来源的DC均表达活化分子(CD86、CD80)，并刺激T细胞增殖。大量的定量RT-PCR研究被用来研究不同培养系统产生的cKit+SCA1+细胞基因表达谱的差异，以及ES来源的细胞和iPS来源的细胞之间的差异。我们的结论是，使用基质条件培养液的无饲养层系统可以有效地从iPS细胞生成HPC和功能树突状细胞，并且所生成的细胞具有与ES细胞相似的基因表达谱。
Patient-specific therapeutic cells derived from induced pluripotent stem (iPS) cells may bypass the ethical issues associated with embryonic stem (ES) cells and avoid potential immunological reactions associated with allogenic transplantation. It is critical, for the ultimate clinical applicability of iPS cell-derived therapies, to establish feeder-free cultures that ensure efficient differentiation of iPS cells into therapeutic progenitors. It is also necessary to understand if iPS cell-derived progenitors differ from those derived from ES cells. In this study, we compared the efficiency of three different feeder-free cultures for differentiating mouse iPS cells into ckit+sca1+ hematopoietic progenitor cells (HPCs) and compared how differentiation and functionality varies between ES and iPS cells. Our results indicated that both iPS and ES cells can be efficiently differentiated into HPCs in suspension cultures supplemented with secretion factors from mouse bone marrow stromal cells (OP9-DL1 conditioned medium). The functionality of these cells was demonstrated by differentiation into CD11c+ dendritic cells (DCs). Both ES and iPS-derived DCs expressed activation molecules (CD86, CD80) in response to LPS stimulation and stimulated T cell proliferation in a mixed lymphocyte reaction (MLR). Extensive quantitative RT-PCR studies were used to study the differences in gene expression profiles of ckit+sca1+ cells generated from the various culture systems as well as differences between ES-derived and iPS-derived cells. We conclude that a feeder-free system using stromal conditioned medium can efficiently generate HPCs as well as functional DCs from iPS cells and the generated cells have similar gene expression profile as those from ES cells.
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影响因子：
5.3
作者：
KELLER, G;KENNEDY, M;WILES, MV
通讯作者：
WILES, MV
影响因子：
30.5
作者：
Schmitt, TM;de Pooter, RF;Zúñiga-Pflücker, JC
通讯作者：
Zúñiga-Pflücker, JC
影响因子：
5.7
作者：
Matsuyoshi, H;Hirata, S;Senju, S
通讯作者：
Senju, S
影响因子：
4.4
作者：
Slukvin, Igor I.;Vodyanik, Maxim A.;Choi, Kyung-Dal
通讯作者：
Choi, Kyung-Dal
影响因子：
4.4
作者：
Matsunaga, Yusuke;Fukuma, Daiki;Senju, Satoru
通讯作者：
Senju, Satoru