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一种高效的人胚胎干细胞早期造血分化和功能筛选模型的建立

, PP. 762-769

Keywords: 人胚胎干细胞,造血分化,功能筛选

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Abstract:

人胚胎干细胞具有分化为造血前体细胞及各种功能血细胞的潜能.但是,目前的分化筛选模型定向诱导分化效率很低,分化程序复杂且较难建立,分化的细胞功能存在缺陷,这些严重影响了对人胚胎干细胞造血分化和功能成熟分子机理的了解.所以,迫切需要建立一种简单且方便进行基因操作的高效的造血分化和功能筛选模型,用于揭示分化过程中动态的分子信号和实时调控机理.本研究中,采用分步法在不依赖血清及基质细胞的条件下,建立了一种高效分化产生造血前体细胞的方法,CD34+CD31+造血前体细胞的比例可达到25%.产生的造血前体细胞可以进一步诱导分化为红系、粒系及粒-巨噬系集落.此外,携带针对T基因干扰shRNA的慢病毒可高效感染人胚胎干细胞,实现高效干扰,调控造血分化效率.本研究建立了一种高效分化产生造血前体细胞的方法,同时表明此模型可以有效与shRNA文库结合,筛选调控人胚胎干细胞造血分化的关键基因,为揭示人胚胎干细胞造血分化机理,以及设计更加完善的分化体系高效获得功能成熟的血细胞用于药物筛选及细胞治疗打下了基础.

References

[1]  1 Thomson J A, Itskovitz-Eldor J, Shapiro S S, et al. Embryonic stem cell lines derived from human blastocysts. Science, 1998, 282: 1145-1147
[2]  2 Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007, 131: 861-872
[3]  3 Kaufman D S. Toward clinical therapies using hematopoietic cells derived from human pluripotent stem cells. Blood, 2009, 114: 3513-3523
[4]  4 Ye Z, Zhan H, Mali P, et al. Human-induced pluripotent stem cells from blood cells of healthy donors and patients with acquired blood disorders. Blood, 2009, 114: 5473-5480
[5]  5 Kaufman D S, Hanson E T, Lewis R L, et al. Hematopoietic colony-forming cells derived from human embryonic stem cells. Proc Natl Acad Sci USA, 2001, 98: 10716-10721
[6]  6 Lu S J, Feng Q, Park J S, et al. Biologic properties and enucleation of red blood cells from human embryonic stem cells. Blood, 2008, 112: 4475-4484
[7]  7 Ma F, Ebihara Y, Umeda K, et al. Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis. Proc Natl Acad Sci USA, 2008, 105: 13087-13092
[8]  8 Takayama N, Nishikii H, Usui J, et al. Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. Blood, 2008, 111: 5298-5306
[9]  9 Takayama N, Eto K. In vitro generation of megakaryocytes and platelets from human embryonic stem cells and induced pluripotent stem cells. Methods Mol Biol, 2012, 788: 205-217
[10]  10 Choi K D, Vodyanik M A, Slukvin I I. Generation of mature human myelomonocytic cells through expansion and differentiation of pluripotent stem cell-derived lin-CD34+CD43+CD45+ progenitors. J Clin Invest, 2009, 119: 2818-2829
[11]  11 Knorr D A, Kaufman D S. Pluripotent stem cell-derived natural killer cells for cancer therapy. Transl Res, 2010, 156: 147-154
[12]  12 Timmermans F, Velghe I, Vanwalleghem L, et al. Generation of T cells from human embryonic stem cell-derived hematopoietic zones. J Immunol, 2009, 182: 6879-6888
[13]  13 Narayan A D, Chase J L, Lewis R L, et al. Human embryonic stem cell-derived hematopoietic cells are capable of engrafting primary as well as secondary fetal sheep recipients. Blood, 2006, 107: 2180-2183
[14]  14 Wang L, Menendez P, Shojaei F, et al. Generation of hematopoietic repopulating cells from human embryonic stem cells independent of ectopic HOXB4 expression. J Exp Med, 2005, 201: 1603-1614
[15]  15 Hexum M K, Tian X, Kaufman D S. In vivo evaluation of putative hematopoietic stem cells derived from human pluripotent stem cells. Methods Mol Biol, 2011, 767: 433-447
[16]  16 Keller G. Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev, 2005, 19: 1129-1155
[17]  17 Dravid G G, Crooks G M. The challenges and promises of blood engineered from human pluripotent stem cells. Adv Drug Deliv Rev, 2011, 63: 331-341
[18]  18 Wang C, Tang X, Sun X, et al. TGFbeta inhibition enhances the generation of hematopoietic progenitors from human ES cell-derived hemogenic endothelial cells using a stepwise strategy. Cell Res, 2012, 22: 194-207
[19]  19 Real P J, Ligero G, Ayllon V, et al. SCL/TAL1 regulates hematopoietic specification from human embryonic stem cells. Mol Ther, 2012, 20: 1443-1453
[20]  20 Vodyanik M A, Bork J A, Thomson J A, et al. Human embryonic stem cell-derived CD34+ cells: efficient production in the coculture with OP9 stromal cells and analysis of lymphohematopoietic potential. Blood, 2005, 105: 617-626
[21]  21 Ledran M H, Krassowska A, Armstrong L, et al. Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches. Cell Stem Cell, 2008, 3: 85-98
[22]  22 Chadwick K, Wang L, Li L, et al. Cytokines and BMP-4 promote hematopoietic differentiation of human embryonic stem cells. Blood, 2003, 102: 906-915
[23]  23 Ng E S, Davis R P, Azzola L, et al. Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation. Blood, 2005, 106: 1601-1603
[24]  24 Yu C, Liu Y, Miao Z, et al. Retinoic acid enhances the generation of hematopoietic progenitors from human embryonic stem cell-derived hemato-vascular precursors. Blood, 2010, 116: 4786-4794
[25]  25 Moreno-Gimeno I, Ledran M H, Lako M. Hematopoietic differentiation from human ESCs as a model for developmental studies and future clinical translations. Invited review following the FEBS Anniversary Prize received on 5 July 2009 at the 34th FEBS Congress in Prague. FEBS J, 2010, 277: 5014-5025

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