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摘要：胚胎分割是哺乳动物胚胎工程的重要组成部分。为细胞和胚胎的相关研究提供有力手段,是增加胚胎数目和生产同卵双胎或多胎,扩大优良畜种后代的一个有效方法。虽然胚胎分割还有许多尚未解决的问题,但其已表现出了广阔的发展前景。 相似文献
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本文介绍了禽类胚胎操作技术方面的概况，对受精卵收集及体外受精、早期胚胎体外培养、人造材料蛋壳培养胚胎、代用蛋壳培养胚胎、嵌合体鸡的生产、转基因鸡生产等的进展及发展进行了综述。 相似文献
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有研究发现,体外生产的胚胎染色体异常的发生率高于体内胚胎。Viuff等人在一项对牛体外胚胎染色体异常的研究中发现,体外生产的牛囊胚的混倍率为72%,而体内生产的囊胚混倍率仅为25%。这说明,胚胎生产的操作过程、培养液成分和其他的一些环境因素可能是引起胚胎发生染色体异常的 相似文献
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猪体外胚胎生产的各个技术环节发展较为成熟,但多偏向于研究,少有推广应用。随着我国家畜资源保护与利用的开展,猪体外胚胎保种势在必行。为向相关工作者提供一个完整高效的生产应用体系,综合分析了包括体外成熟、体外受精和体外培养等方面的技术要点,并总结有关猪体外胚胎生产各关键技术环节的发展与应用现状,以期为进一步保障我国猪产业经济的发展,提供理论参考。 相似文献
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胚胎工程技术在畜牧业上的应用 总被引：1，自引：0，他引：1
胚胎工程或叫胚胎技术 ,是以胚胎移植为基础 ,以转基因动物为中心 ,通过人工操作胚胎 ,定向改变动物性状使之为人类造福的一系列生物技术的总称。主要包括 :胚胎移植、体外胚胎生产 (体外成熟、体外受精和胚胎培养 )、胚胎冷冻、胚胎分割、动物克隆、胚胎性别鉴定、胚胎嵌合、显微受精和转基因动物技术等。1 目前可以在畜牧业上应用的技术胚胎移植、体外胚胎生产 (体外成熟、体外受精和胚胎培养、胚胎冷冻 )、胚胎分割等胚胎工程技术在国外已经商业化 ,牛和羊的胚胎移植技术在我省的畜牧业上的应用前景最大。要开展胚胎移植技术 ,还需要开… 相似文献
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随着商业性胚胎移植技术的推广应用和动物胚胎工程技术的深入研究,对胚胎和卵母细胞的需求量日益增加,单靠超排技术获得的胚胎或卵母细胞已远远不能满足科研和生产的需要,因而推动了卵母细胞体外成熟（IVM）、体外受精（IVF）及体外胚胎生产（IVP）等胚胎工程技术的发展。牛体外受精技术日趋成熟,体外胚胎生产已经开始走向生产应用,但山羊体外胚胎的生产效率远落后于牛,且质量不稳定。优化培养条件是提高山羊胚胎生产效率和胚胎质量的关键。文献报道,将胶质细胞源性神经营养因子（GDNF）、白血病抑制因子（LIF）、干细胞因子（SCF）用于绵羊、小鼠等动物的细胞培养研究取得了较理想的效果,但未见将3种成分用于山羊的报道。 相似文献
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应用体外受精技术,可以有效地利用屠宰的青年母猪卵巢中的未成熟卵,大量、廉价的实验室生产胚胎。这对加速我国良种猪群的繁殖是一项行之有效的胚胎工程技术。借鉴国内外实验室生产猪胚胎的经验及根据实验室的实践,文章较详细地叙述了体外受精猪胚胎的生产过程,包括卵母细胞的获得、培养、精子获能、体外受精以及受精后的体外培养、冷冻保存与移植等。 相似文献
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本文对哺乳动物胚胎细胞体细胞核移植克隆技术中显微操作的基本环节,以及细胞周期阶段对细胞核移植克隆胚胎效率影响的研究进行综述。 相似文献
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胚胎干细胞是未分化的具有增殖和自我更新能力的细胞,并且能分化成所有类型的体细胞以及生殖细胞。它们提供了早期胚胎分化的体外模型,也是基因操作的重要靶细胞。禽类多能性干细胞培养最重要的应用领域是以干细胞体外遗传修饰、鉴定为技术平台的家禽转基因技术。通过此技术对禽类基因进行遗传修饰与操作,在胚胎发育基础研究、转基因禽类生产及家禽育种等方面有巨大的应用前景。但是禽类多能性干细胞培养的许多基本问题仍亟待解决,如探索其建系的培养条件、揭示其维持多能性和增殖能力的分子机制等。文章综述了禽类多能性干细胞的分离方法、体外分化能力、嵌合体形成以及基因修饰方面的研究进展及目前的研究局限。 相似文献
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P Blackshear J Mahler L M Bennett K A McAllister D Forsythe B J Davis 《Veterinary pathology》1999,36(5):457-460
Chimeric mice often are created through the genetic manipulation of the mouse embryo in the process of developing animal models of disease. These mice have variable percentages of their somatic and germ cells derived from the donor embryonic stem cells and host blastocysts. In the development of mouse models deficient in the breast cancer susceptibility gene 2 (Brca2) or the 70-kd heat shock protein (Hsp70-2), 3-4-week-old chimeras developed single or multiple masses composed of both well-differentiated and poorly differentiated tissues derived from all three germ layers. These cases of extragonadal teratocarcinoma, a rarely reported tumor, may be related to the genetic predisposition of the 129/Ola mouse strain used to generate the embryonic stem cells. 相似文献
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位点特异性重组系统已经成为基因组操作的强有力工具。来自链霉菌噬菌体φC31的整合酶是丝氨酸重组酶家族的一员,可催化2个不同的识别位点attB和attP间发生有效的单向性重组。总结了φC31整合酶可能的重组机制以及在人类细胞、小鼠胚胎干细胞、果蝇和植物等真核生物基因组操作上的应用,并对该系统的优点和存在问题进行了分析。φC31整合酶系统有望成为高等生物基因功能分析、转基因标记删除、生物合成等研究领域最强有力的手段之一,为此也简要介绍了家蚕个体水平上φC31整合酶介导的盒式交换的研究进展,期望能为家蚕基因组的操作技术研究提供一些借鉴。 相似文献
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Recent Progress in Embryonic Stem Cell Research and Its Application in Domestic Species 总被引：1，自引：0，他引：1
TAL Brevini S Antonini G Pennarossa F Gandolfi 《Reproduction in domestic animals》2008,43(S2):193-199
Many reports described cell lines derived in domestic species, which presented several important features typical of embryonic stem cells (ESCs). Such features unfortunately did not include the capacity to generate germ-line chimeras, therefore limiting the possibility to use these cells as tools for the genetic manipulation. However, farm animal ESCs may still be useful for the generation of transgenic animals as usually have a self-renewal capacity more prolonged than normal primary cultures thus increasing the possibility to transform and select cells to be used as nucleus donors in cloning procedures. Farm animal ESCs may also be an excellent experimental model in pre-clinical trials, assessing the feasibility of cell therapy because of the close morphological and physiological resemblance to humans of species like the pig. However, the persistent lack of standard methods for the derivation, maintenance and characterization of ESCs in domestic species stimulated the search for alternatives. Embryonic germ cells may represent such an alternative. Indeed, these cells showed a higher plasticity than ESCs as contributed to embryonic development forming chimeric newborns but, as for ESCs, standardization is still far away and efficiency is very low. Recent results indicated spermatogonial stem cells as possible tools for germ-line genetic modifications with some proof of principle results already achieved. But, a real break through could arrive from the multipotent germ-line stem cells, virtually equivalent to ESC, derived from newborn and adult mouse testis. 相似文献
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1. The present study was carried out to determine whether primordial germ cells isolated from embryonic blood can enter the bloodstream and successfully migrate to the germinal ridges of recipient embryos after transfer to stage X blastoderms, and also whether they can differentiate into blood cells, as is suggested in mice. 2. Primordial germ cells were transfected in vitro by lipofection and then transferred to stage X blastoderms. The introduced GFP gene was efficiently expressed in the gonads of 6-d incubated embryos. 3. Freshly collected primordial germ cells were transferred to stage X blastoderms. The fate of the transferred primordial germ cells was traced by detecting the single nucleotide polymorphism in the D-loop region of the mitochondrial DNA in White Leghorn and Barred Plymouth Rock chickens used in this study. The transferred donor primordial germ cell-derived cells were detected in the gonads, but not in the blood cells, of 17-d incubated embryos by PCR. 4. This procedure for primordial germ cell manipulation could provide a novel method of producing germline chimaeric chickens. 5. In conclusion, our findings indicate that primordial germ cells isolated from embryonic blood can migrate to the germinal ridges of recipient embryos after being transferred to stage X blastoderms. Although these transferred primordial germ cells differentiated into germ cells, no differentiation into blood cells was observed. 相似文献
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Porcine embryonic fibroblasts (PEFs) are widely used as donor cells for somatic cell nuclear transfer (SCNT) in pigs. Transfection of PEFs with exogenous DNA is essential for producing genetically modified (GM; transgenic or knockout) pigs via SCNT. In this case, selectable markers are strictly required selecting and enriching stably transfected cells. The most frequently used selective drug for this purpose is a neomycin analogue (G418/geneticin); neo has been widely used as a selectable marker gene in the genomic manipulation of pigs. However, little is known about optimal concentrations of other selection drugs. This often hampers functional analysis of the porcine genome and development of individual GM pigs. This study explores the optimal concentrations of selective drugs, other than neomycin, that can be used for the selection of transfected PEFs. Porcine embryonic fibroblasts were incubated in media containing different concentrations of drugs for up to 10 days, to determine the optimal drug concentrations fatal for PEFs. The following concentrations were found to be optimal selective concentrations for use with PEFs: G418/geneticin, 400 μg/ml; blasticidin S, 8 μg/ml; hygromycin B, 40 μg/ml; puromycin, 2 μg/ml; and zeocin, 800 μg/ml. Repeated transfections with plasmids carrying selectable markers resulted in the generation of multidrug-resistant swine transfectants. Furthermore, these markers were found to be independent. The present information will be useful for the production of SCNT-mediated GM piglets that express multiple transgenes. 相似文献
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I Dobrinski 《Reproduction in domestic animals》2008,43(S2):288-294
Transplantation of male germ line stem cells from a donor animal to the testes of an infertile recipient was first described in 1994. Donor germ cells colonize the recipient's testis and produce donor-derived sperm, such that the recipient male can distribute the genetic material of the germ cell donor. Germ cell transplantation represents a functional reconstitution assay for male germ line stem cells and as such has vastly increased our ability to study the biology of stem cells in the testis and define phenotypes of infertility. First developed in rodents, the technique has now been used in a number of animal species, including domestic mammals, chicken and fish. There are three major applications for this technology in animals: first, to study fundamental aspects of male germ line stem cell biology and male fertility; second, to preserve the reproductive potential of genetically valuable individuals by male germ cell transplantation within or between species; third, to produce transgenic sperm by genetic manipulation of isolated germ line stem cells and subsequent transplantation. Transgenesis through the male germ line has tremendous potential in species in which embryonic stem cells are not available and somatic cell nuclear transfer has limited success. Therefore, transplantation of male germ cells is a uniquely valuable approach for the study, preservation and manipulation of male fertility in animals. 相似文献
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Xi Y Nada Y Soh T Fujihara N Hattori MA 《The Journal of reproduction and development》2003,49(3):213-219
The present study was performed to develop a culture system for feather keratinocyte stem cells to enable the genetic manipulation of endangered avian species. The feather follicle cells were isolated from growing feathers of adult White Leghorn chicken. Leukemia inhibitory factor (LIF) was used to maintain the characterization of the keratinocyte colony-forming cells (KCFCs). The EGFPN1 plasmid DNA retroviral vector was used to deliver Green Fluorescent Protein (GFP) gene, which was introduced to the KCFCs by lipofection. After removal of the fibroblast-like cells, the feather KCFCs attached to the substrate within 24 h of seeding. The cells continued to proliferate for at least 30 days in the presence of LIF. The cell-adhesion molecules such as integrin beta1 and CD49c were immunocytochemically positive in the cells. The KCFCs differentiated into barbular cells and pennaceous feather vane in the LIF-free medium. The GFP gene-transfected KCFCs stably expressed GFP. The present results indicate that the KCFCs derived from feather follicles are closely related to multipotent stem cells. In addition, gene manipulation of such stem cells may be useful for the production of chimera in avian species. 相似文献
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鸡胚胎干细胞是一种多能性干细胞,从X期胚盘分离胚盘细胞或早期鸡胚的生殖嵴分离原始生殖细胞,经体外长期抑制分化培养可得到鸡胚胎干细胞。为维持细胞在培养过程中的未分化状态,需要采用饲养层细胞培养,同时设计合理的培养液配方并添加多种抑制分化或促进增殖的细胞因子。通过碱性磷酸酶活性检测、胚胎表面特异性抗原检测、分化试验及嵌合体试验等方法,可对鸡胚胎干细胞进行准确鉴定。文章主要就鸡胚胎干细胞的分离、培养与鉴定方法的研究进展及其应用前景进行简要综述,为进一步发展更高效的鸡胚胎干细胞培养体系并应用于生产实践提供一定的借鉴。 相似文献