拟胚体对成体细胞参与胚胎嵌合的影响

曹鸿国,李运生,曲秀霞,邓宏魁

中国农学通报. 2008, 24(9): 25-30

中国农学通报 ›› 2008, Vol. 24 ›› Issue (9) : 25-30. DOI: 10.11924/j.issn.1000-6850.2008-0483-20086151
畜牧 动物医学 蚕 蜂

拟胚体对成体细胞参与胚胎嵌合的影响

  • 曹鸿国,李运生,曲秀霞,邓宏魁
作者信息 +

The Effect on Chimeric Embryo Construction from Fibroblasts and Early
Embryos about Mouse Embryoid Body

  • Cao Hongguo, Li Yunsheng, Qu Xiuxia, Deng Hongkui
Author information +
History +

摘要

试验尝试构建成体细胞同小鼠早期胚胎的嵌合共生胚胎。取成年人皮肤组织的成纤维细胞，慢病毒转染皮肤成纤维细胞标记EGFP荧光蛋白，同小鼠早期8-细胞胚胎进行嵌合。结果显示慢病毒高效标记人皮肤成纤维细胞表达EGFP荧光蛋白，通过皮肤成纤维细胞与小鼠胚胎干细胞形成的拟胚体环境作用后，皮肤成纤维细胞成功与小鼠胚胎形成嵌合胚，嵌合囊胚形成率为38.08%，表达EGFP荧光蛋白的皮肤成纤维细胞能够嵌合到小鼠胚胎的不同部位。嵌合胚在胚胎干细胞分离培养环境下进行培养，嵌合到小鼠内细胞团的皮肤成纤维细胞参与小鼠内细胞团的组成，参与小鼠内细胞团组成的胚胎占嵌合胚比率为1.74 %。小鼠胚胎干细胞拟胚体环境作用可以成功介导人皮肤成纤维细胞同小鼠早期胚胎形成嵌合共生体系。

Abstract

Chimeric embryos from mouse embryos and human fibroblasts were constructed. The fibroblasts from adult foreskin were isolated, cultured and transfected EGFP with lentivirus vectors, and then chimeric embryos were constructed with mouse embryos and fibroblasts. The results showed that human fibroblasts efficiently expressed EGFP after transfected with lentivirus vectors. 38.08 % chimeric embryos were constructed successfully with mouse embryos and EGFP fibroblasts which were from embryoid bodies of mouse embryonic stem cells and EGFP fibroblasts. EGFP fibroblasts were taken part in different ingredient of chimeric embryos. Under the circumstance of embryonic stem cell isolation and culture, EGFP fibroblasts in 1.74 % chimeric embryos became the member of inner cell masses on feeder cells. Human fibroblasts can become chimeric embryos with mouse embryos by the microenvironment of mouse embryoid bodies.

关键词

拟胚体；成纤维细胞；胚胎；嵌合

Key words

embryoid body;fibroblast;embryo;chimera

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曹鸿国,李运生,曲秀霞,邓宏魁. 拟胚体对成体细胞参与胚胎嵌合的影响. 中国农学通报. 2008, 24(9): 25-30 https://doi.org/10.11924/j.issn.1000-6850.2008-0483-20086151
Cao Hongguo, Li Yunsheng, Qu Xiuxia, Deng Hongkui. The Effect on Chimeric Embryo Construction from Fibroblasts and Early
Embryos about Mouse Embryoid Body. Chinese Agricultural Science Bulletin. 2008, 24(9): 25-30 https://doi.org/10.11924/j.issn.1000-6850.2008-0483-20086151

参考文献

[1]Zeng F, Chen M J, Baldwin D A, et al. Multiorgan engraftment and differentiation of human cord blood CD34+ Lin-cells in goats assessed by gene expression profiling. Proc Natl Acad Sci USA, 2006, 103(20):7801-806.
[2]Kulesa P M, Kasemeier-Kulesa J C, Teddy J M, et al. Reprogramming metastatic melanoma cells to assume a neural crest cell-like phenotype in an embryonic microenvironment. Proc Natl Acad Sci USA, 2006, 103(10): 3752-3757.
[3]Chen F G, Zhang W J, Bi D, et al. Clonal analysis of nestin(-) vimentin(+) multipotent fibroblasts isolated from human dermis. J Cell Sci, 2007, 120 (Pt16):2875-2883.
[4]Sterin-Borda L, Furlan C, Orman B, et al. Differential regulation on human skin fibroblast by alpha1 adrenergic receptor subtypes. Biochem Pharmacol, 2007, 74(9):1401-1142.
[5]Meng G L, Zur Nieden N I, Liu S Y, et al. Properties of murine embryonic stem cells maintained on human foreskin fibroblasts without LIF. Mol Reprod Dev, 2008, 75(4): 614-622.
[6]Amit M, Margulets V, Segev H, et al. Human feeder layers for human embryonic stem cells. Biol Reprod, 2003, 68(6): 2150-2156.
[7]Inzunza J, Gertow K, Str?觟mberg M A, et al. Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells. Stem Cells, 2005, 23(4): 544-549.
[8]Valbuena D, Galán A, Sánchez E, et al. Derivation and characterization of three new Spanish human embryonic stem cell lines (VAL-3 -4 -5) on human feeder and in serum-free conditions. Reprod Biomed, 2006, 13(6): 875-886.
[9]Nieto A, Cabrera C M, Catalina P, et al. Effect of mitomycin-C on human foreskin fibroblasts used as feeders in human embryonic stem cells: immunocytochemistry MIB1 score and DNA ploidy and apoptosis evaluated by flow cytometry. Cell Biol Int, 2007, 31(3): 269-278.
[10]Yu J, Vodyanik M A, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007, 318(5858): 1917-1920.
[11]Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007, 131(5): 861-872.
[12]LinksGaly A, Roncarolo M G, Thrasher A J. Development of lentiviral gene therapy for Wiskott Aldrich syndrome. Expert Opin Biol Ther, 2008, 8(2): 181-190.
[13]Tannemaat M R, Boer G J, Verhaagen J, et al. Genetic modification of human sural nerve segments by a lentiviral vector encoding nerve growth factor. Neurosurgery, 2007, 61(6): 1286-1294.
[14]Kim M, Carman C V, Yang W, et al. The primacy of affinity over clustering in regulation of adhesiveness of the integrin {alpha}L{beta}2. J Cell Biol, 2004, 167(6): 1241-1253.
[15]Luo B H, Carman C V, Springer T A. Structural basis of integrin regulation and signaling. Annu Rev Immunol, 2007, 25: 619-647.
[16]Shimaoka M, Takagi J, Springer T A. Conformational regulation of integrin structure and function. Annu Rev Biophys Biomol Struct, 2002, 31: 485-516.
[17]Illera M J, Lorenzo P L, Gui Y T, et al. A role for alphavbeta3 integrin during implantation in the rabbit model. Biol Reprod, 2003, 68(3): 766-771.
[18]Tian X, Morris J K, Linehan J L, et al. Cytokine requirements differ for stroma and embryoid body-mediated hematopoiesis from human embryonic stem cells. Exp Hematol, 2004, 32(10): 1000-1009.
[19]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(19): 10716-10721.
[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(2): 617-626.
[21]Qiu C, Hanson E, Olivier E, et al. Differentiation of human embryonic stem cells into hematopoietic cells by coculture with human fetal liver cells recapitulates the globin switch that occurs early in development. Exp Hematol, 2005, 33(12): 1450-1458.
[22]Fu J R, Liu W L, Zhou Y F, et al. Expansive effects of aorta- gonad-mesonephros-derived stromal cells on hematopoietic stem cells from embryonic stem cells. Chin Med J (Engl), 2005, 118(23): 1979-1986.
[23]Wang J, Zhao H P, Lin G, et al. In vitro hematopoietic differentiation of human embryonic stem cells induced by co-culture with human bone marrow stromal cells and low dose cytokines. Cell Biol Int, 2005, 29(8): 654-561.
[24]Chen S, Zhang Q, Wu X, Schultz P G, et al. Dedifferentiation of lineage-committed cells by a small molecule. J Am Chem Soc, 2004, 126(2): 410-411.
[25]Gage F H. Mammalian neural stem cells. Science, 2000, 287(25): 1433-1438.
[26]Davis M E, Hsieh P C, Takahashi T, et al. Local myocardial insulin-like growth factor 1 (IGF-1) delivery with biotinylated peptide nanofibers improves cell therapy for myocardial infarction. Proc Natl Acad Sci USA. 2006, 103(21): 8155-8160.
[27]Fu X, Sun X, Li X, et al. Dedifferentiation of epidermal cells to stem cells in vivo. Lancet, 2001, 358(9287): 1067-1068.


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