Gene expression profile and functionality of ESC-derived Lin-ckit+Sca-1+ cells are distinct from Lin-ckit+Sca-1+ cells isolated from fetal liver or bone marrow.
Gene expression profile and functionality of ESC-derived Lin-ckit+Sca-1+ cells are distinct from Lin-ckit+Sca-1+ cells isolated from fetal liver or bone marrow.
复制标题
DOI：
10.1371/journal.pone.0051944
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Roy K
中科院分区：
文献类型：
--
作者：
Fernandez I;Fridley KM;Arasappan D;Ambler RV;Tucker PW;Roy K
基于体外生物反应器的培养物正在被广泛研究用于从胚胎干细胞（ESC）大规模生产分化细胞。然而，目前尚不清楚体外ESC衍生的祖细胞是否具有与体内对应物相似的基因表达谱和功能。这对于确定ESC衍生细胞作为成人分离细胞的替代品用于临床治疗的有效性至关重要。在这项研究中，我们比较了Lin-ckit+Sca-1+（LKS）细胞在体外产生的小鼠胚胎干细胞使用静态或生物反应器为基础的培养物，与从小鼠胎肝（FL）或骨髓（BM）分离的天然LKS细胞的基因表达谱。我们发现，在体外产生的LKS细胞更类似于FL-比BM LKS细胞的基因表达。此外，当与来自生物反应器培养物的细胞相比时，静态培养物来源的LKS细胞相对于两种体内LKS群体显示出较少的差异表达基因。总的来说，造血基因的表达在ESC衍生的LKS细胞中比来自BM和FL的细胞低，而非造血基因的水平上调。为了确定这些分子特征是否与功能相关，我们评估了ESC衍生的LKS细胞的体外造血分化和集落形成（CFU测定）。尽管静态培养产生的细胞未能形成任何集落，但它们确实分化为CD 11 c+和B220+细胞，表明具有一定的造血潜力。相反，生物反应器衍生的LKS细胞在相同条件下分化时不能产生任何B220+或CD 11 c+细胞，也不形成集落，表明这些细胞不是造血祖细胞。我们的结论是，在体外培养条件下显着影响转录组和功能的ESC衍生的LKS细胞，虽然在体外分化的LKS细胞是谱系阴性，并表达ckit和Sca-1，这些细胞，特别是那些从动态培养，是显着不同的天然细胞相同的表型。
In vitro bioreactor-based cultures are being extensively investigated for large-scale production of differentiated cells from embryonic stem cells (ESCs). However, it is unclear whether in vitro ESC-derived progenitors have similar gene expression profiles and functionalities as their in vivo counterparts. This is crucial in establishing the validity of ESC-derived cells as replacements for adult-isolated cells for clinical therapies. In this study, we compared the gene expression profiles of Lin-ckit+Sca-1+ (LKS) cells generated in vitro from mouse ESCs using either static or bioreactor-based cultures, with that of native LKS cells isolated from mouse fetal liver (FL) or bone marrow (BM). We found that in vitro-generated LKS cells were more similar to FL- than to BM LKS cells in gene expression. Further, when compared to cells derived from bioreactor cultures, static culture-derived LKS cells showed fewer differentially expressed genes relative to both in vivo LKS populations. Overall, the expression of hematopoietic genes was lower in ESC-derived LKS cells compared to cells from BM and FL, while the levels of non-hematopoietic genes were up-regulated. In order to determine if these molecular profiles correlated with functionality, we evaluated ESC-derived LKS cells for in vitro hematopoietic-differentiation and colony formation (CFU assay). Although static culture-generated cells failed to form any colonies, they did differentiate into CD11c+ and B220+ cells indicating some hematopoietic potential. In contrast, bioreactor-derived LKS cells, when differentiated under the same conditions failed to produce any B220+ or CD11c+ cells and did not form colonies, indicating that these cells are not hematopoietic progenitors. We conclude that in vitro culture conditions significantly affect the transcriptome and functionality of ESC-derived LKS cells and although in vitro differentiated LKS cells were lineage negative and expressed both ckit and Sca-1, these cells, especially those obtained from dynamic cultures, are significantly different from native cells of the same phenotype.
登录
查看更多内容
小鼠骨髓中多能间充质干细胞的前瞻性鉴定、分离和系统移植。
DOI：
10.1084/jem.20091046
发表时间：
2009-10-26
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Morikawa S;Mabuchi Y;Kubota Y;Nagai Y;Niibe K;Hiratsu E;Suzuki S;Miyauchi-Hara C;Nagoshi N;Sunabori T;Shimmura S;Miyawaki A;Nakagawa T;Suda T;Okano H;Matsuzaki Y
通讯作者：
Matsuzaki Y
影响因子：
20.3
作者：
Mikkola, HKA;Fujiwara, Y;Orkin, SH
通讯作者：
Orkin, SH
影响因子：
64.5
作者：
Kiel, MJ;Yilmaz, ÖH;Morrison, SJ
通讯作者：
Morrison, SJ
影响因子：
4.1
作者：
Fridley, Krista M.;Fernandez, Irina;Roy, Krishnendu
通讯作者：
Roy, Krishnendu
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y