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1. 发明申请

US20050193435A1 Prolonged culturing of avian (chicken) primordial germ cells (PGCs) using specific growth factors, use thereof to produce chimeric avians (chickens) 审中-公开
标题翻译：使用特定生长因子长时间培养禽类（鸡）原始生殖细胞（PGCs），利用其生产嵌合禽鸟（鸡）
公开(公告)号：US20050193435A1
公开(公告)日：2005-09-01
申请号：US10934751
申请日：2004-09-03
申请人： James Robl , Steven Stice , D. Jerry , F. Leon , Catherine Blackwell , Xiu Gao
发明人： James Robl , Steven Stice , D. Jerry , F. Leon , Catherine Blackwell , Xiu Gao
IPC分类号： A01K67/027 , C12N5/074 , C12N5/06
CPC分类号： A01K67/0275 , A01K67/0271 , A01K2217/05 , A01K2227/30 , C12N5/0611 , C12N2501/105 , C12N2501/115 , C12N2501/125 , C12N2501/235
摘要： A culture system for maintaining avian PGCs for long periods in tissue culture is provided. This culture system uses LIF, bFGF, IGF and SCF. The resultant PGCs are useful for the production of transgenic and chimeric avians, in particular, chickens or turkeys.
摘要翻译：提供了一种用于在组织培养中长期维持禽类PGC的培养系统。该文化系统使用LIF，bFGF，IGF和SCF。所得的PGC可用于生产转基因和嵌合鸟类，特别是鸡或火鸡。

2. 发明授权

US07547818B2 Metaphase donor cells for effective nuclear reprogramming in mammals 失效
标题翻译：中期供体细胞用于哺乳动物的有效核重编程
公开(公告)号：US07547818B2
公开(公告)日：2009-06-16
申请号：US10809738
申请日：2004-03-25
申请人： Steven Stice
发明人： Steven Stice
IPC分类号： C12N15/00
CPC分类号： C12N15/873
摘要： The present invention provides methods of producing a clone non-human mammalian nuclear transfer (NT) embryo and methods for producing a cloned non-human mammal. Embodiments of the methods include introducing doner genetic material into a metaphase I oocyte; introducing donor genetic material into a non-enucleated oocyte; introducing donor genetic material obtained from a donor cell that is a metaphase into an oocyte; introducing donor genetic material into an oocyte, and naturally activating the oocyte or the NT embryo; and introducing donor genetic material obtained from a donor cell that is at late G1 phase into anoocyte.
摘要翻译：本发明提供了产生克隆非人哺乳动物核转移（NT）胚胎的方法和用于产生克隆的非人哺乳动物的方法。方法的实施方案包括将内含子遗传物质引入中期卵母细胞; 将供体遗传物质引入非去核卵母细胞; 将从中期的供体细胞获得的供体遗传物质引入卵母细胞; 将供体遗传物质引入卵母细胞，并自然激活卵母细胞或NT胚胎; 并将从晚期G1期的供体细胞获得的供体遗传物质引入到不孕细胞中。

3. 发明申请

US20050244958A1 Production of avian embryonic germ (EG) cell lines by prolonged culturing of PGCs, use thereof for cloning and chimerization 审中-公开
标题翻译：通过长期培养PGC来生产禽胚胎胚芽（EG）细胞系，用于克隆和嵌合
公开(公告)号：US20050244958A1
公开(公告)日：2005-11-03
申请号：US11081790
申请日：2005-03-09
申请人： F. Ponce de Leon , James Robl , Steven Stice , D. Jerry
发明人： F. Ponce de Leon , James Robl , Steven Stice , D. Jerry
IPC分类号： C12N5/074 , C12N5/06
CPC分类号： C12N5/0611 , C12N2500/44 , C12N2501/105 , C12N2501/115 , C12N2501/125 , C12N2501/235
摘要： A culture system for producing PGCs or EG cells by culturing PGCs for long periods in tissue culture is provided. This culture system uses LIF, bFGF, IGF and SCF. The resultant EG cells are useful for the production of transgenic and chimeric avians, in particular, chickens and turkeys, and also for cloning purposes.
摘要翻译：提供了通过在组织培养中长期培养PGC来生产PGC或EG细胞的培养系统。该文化系统使用LIF，bFGF，IGF和SCF。所产生的EG细胞可用于生产转基因和嵌合鸟类，特别是鸡和火鸡，以及用于克隆目的。

4. 发明申请

US20100184212A1 HUMAN EMBRYONIC STEM CELL DERIVED MESODERM-LIKE EPITHELIUM TRANSITIONS TO MESENCHYMAL PROGENITOR CELLS 审中-公开
标题翻译：人胚胎干细胞衍生的MESODERM型上皮细胞转染到细胞祖细胞
公开(公告)号：US20100184212A1
公开(公告)日：2010-07-22
申请号：US12451720
申请日：2008-05-30
申请人： Steven Stice , Nolan Boyd
发明人： Steven Stice , Nolan Boyd
IPC分类号： C12N5/0735 , C12N5/0789 , C12N5/077
CPC分类号： C12N5/0662 , C12N5/0625 , C12N2501/105 , C12N2501/11 , C12N2501/115 , C12N2501/165 , C12N2506/02
摘要： Human embryonic stem cells (hESC) have the potential to produce all of the cells in the body. They are also able to self-renew indefinitely, sparking the hope they could be used as a source for large scale production of therapeutic cell lines. The present invention relates to a monolayer differentiation culture system that induces hESC (WA09 and BG01) to form epithelial sheets with mesodermal gene expression patterns (BMP4, RUNX1, GAT A4). These E-cadherin+ CD90lovv cells then undergo apparent epithelial-mesenchymal transformation (EMT) for the derivation of mesenchymal progenitor cells (hES-MC) that by flow cytometry are negative for hematopoietic (CD34, CD45 and CD 133) and endothelial (CD31 and CD 146) markers, but positive for markers associated with mesenchymal stem cells (MSC) (CD73, CD90, CD105 and CD166). To determine their functionality, we tested their capacity to produce the three lineages commonly associated with MSC and found they could form osteogenic and chondrogenic, but not adipogenic lineages. The derived hES-MC were able to remodel and contract collagen I lattice constructs to an equivalent degree as keloid fibroblast control cells and were induced to express αSMA when exposed to TGF-β1, but not PDGF-B. This data suggests the derived hES-MC cells are multipotent cells with potential uses in tissue engineering/regenerative medicine and for providing a highly reproducible cell source for adult-like progenitor cells.
摘要翻译：人类胚胎干细胞（hESC）具有产生体内所有细胞的潜力。他们也能够无限期地自我更新，激发了他们希望将其用作大规模生产治疗性细胞系的来源。本发明涉及诱导hESC（WA09和BG01）形成具有中胚层基因表达模式（BMP4，RUNX1，GAT A4）的上皮片的单层分化培养系统。这些E-钙粘蛋白+ CD90lovv细胞然后经历明显的上皮 - 间质转化（EMT），用于衍生间充质祖细胞（hES-MC），其通过流式细胞术对造血（CD34，CD45和CD133）和内皮（CD31和CD 146）标记，但与间充质干细胞（MSC）（CD73，CD90，CD105和CD166）相关标志物阳性。为了确定其功能，我们测试了其产生通常与MSC相关的三个谱系的能力，发现它们可以形成成骨和软骨形成，但不能形成脂肪形成的谱系。衍生的hES-MC能够将胶原I晶格构建体重塑和收缩至瘢痕疙瘩成纤维细胞对照细胞的相当程度，并且当暴露于TGF-β1而不是PDGF-B时被诱导表达αSMA。该数据表明衍生的hES-MC细胞是在组织工程/再生医学中具有潜在用途的多能细胞，并为成年样祖细胞提供高度可重复的细胞来源。

5. 发明申请

US20060294609A1 Production of cloned offspring from cooled carcasses 审中-公开
标题翻译：从冷却的屠体生产克隆的后代
公开(公告)号：US20060294609A1
公开(公告)日：2006-12-28
申请号：US10512153
申请日：2003-04-24
申请人： Steven Stice , John Gibbons , Donald Respess
发明人： Steven Stice , John Gibbons , Donald Respess
IPC分类号： A01K67/027
CPC分类号： C12N15/8771
摘要： Genetic material is derived from animals post-mortem, and used in nuclear transfer processes to produce cloned embryos and live cloned animals having genetic make-ups identical to the post mortem animals. The method has particular applicability to the management and breeding of livestock, to the production of animals having desired genetic traits, and to the integration of those genetic traits into selective breeding operations.
摘要翻译：遗传物质来源于宰后动物，并用于核转移过程中，以产生具有与死后动物相同的遗传构成的克隆胚胎和活的克隆动物。该方法特别适用于牲畜的管理和育种，具有所需遗传性状的动物的生产，以及将这些遗传特性整合到选育育种操作中。

6. 发明申请

US20050250203A1 Embryonic or stem-like cell lines produced by cross species nuclear transplantation 审中-公开
公开(公告)号：US20050250203A1
公开(公告)日：2005-11-10
申请号：US10981137
申请日：2004-11-04
申请人： James Robl , Jose Cibelli , Steven Stice
发明人： James Robl , Jose Cibelli , Steven Stice
IPC分类号： C12N15/09 , A61K35/12 , A61K38/00 , A61K48/00 , A61P1/16 , A61P3/10 , A61P9/00 , A61P13/00 , A61P25/16 , A61P25/28 , A61P31/18 , A61P35/00 , C12N5/10 , C12N15/85 , C12N15/87 , C12N15/877 , C12N5/08
CPC分类号： C12N15/8776 , A61K35/12 , A61K38/00 , C12N15/87 , C12N2517/04
摘要： An improved method of nuclear transfer involving the transplantation of donor cell nuclei into enucleated oocytes of a species different from the donor cell is provided. The resultant nuclear transfer units are useful for the production of isogenic embryonic stem cells, in particular human isogenic embryonic or stem cells. These embryonic or stem-like cells are useful for producing desired differentiated cells and for introduction, removal or modification, of desired genes, e.g., at specific sites of the genome of such cells by homologous recombination. These cells, which may contain a heterologous gene, are especially useful in cell transplantation therapies and for in vitro study of cell differentiation.

7. 发明申请

US20050108785A1 Cloning using donor nuclei from differentiated fetal and adult cells 审中-公开
标题翻译：使用来自分化胎儿和成体细胞的供体核克隆
公开(公告)号：US20050108785A1
公开(公告)日：2005-05-19
申请号：US10932921
申请日：2004-09-02
申请人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. de Leon , D. Jerry
发明人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. de Leon , D. Jerry
IPC分类号： A01K67/02 , A01K67/027 , A61K35/12 , A61P1/18 , A61P3/10 , A61P9/00 , A61P9/04 , A61P13/02 , A61P17/02 , A61P21/00 , A61P25/16 , A61P25/28 , A61P31/18 , A61P35/00 , C12N5/073 , C12N5/10 , C12N15/09 , C12N15/877
CPC分类号： A01K67/0275 , A01K67/027 , A01K2217/05 , A01K2227/101 , A01K2227/106 , A61K35/12 , C12N5/0603 , C12N15/8771 , C12N15/8778 , C12N2510/00 , C12N2517/02 , C12N2517/04
摘要： An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
摘要翻译：提供了涉及将供体分化的细胞核移植到与供体细胞相同物种的去核卵母细胞中的改进的核转移方法。所产生的核转移单位可用于通过胎儿和后代的产生以及用于产生包括人同种异体胚胎或干细胞在内的等基因CICM细胞的基因型和转基因基因型的倍增。通过本方法促进遗传工程或转基因哺乳动物胚胎，胎儿和后代的生产，因为供体核的分化细胞源可以被遗传修饰和克隆繁殖。

8. 发明申请

US20070067861A1 Cloning pigs using donor nuclei from differentiated cells 审中-公开
标题翻译：使用来自分化细胞的供体核克隆猪
公开(公告)号：US20070067861A1
公开(公告)日：2007-03-22
申请号：US11603036
申请日：2006-11-22
申请人： Steven Stice , James Robl , Jose Cibelli , Paul Golueke
发明人： Steven Stice , James Robl , Jose Cibelli , Paul Golueke
IPC分类号： A01K67/027
CPC分类号： C12N15/8778 , A01K67/027 , A01K67/0273 , A01K67/0275 , A01K2217/05 , A01K2227/101 , A61K35/12 , A61K38/00 , C12N5/0603 , C12N15/8771 , C12N2510/00 , C12N2517/02 , C12N2517/04
摘要： An improved method of nuclear transfer involving the transplantation of donor differentiated pig cell nuclei into enucleated pig oocytes is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring. Production of genetically engineered or transgenic pig embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
摘要翻译：提供了涉及将供体分化的猪细胞核移植到去核猪卵母细胞中的改进的核转移方法。所产生的核转移单位可用于通过胎儿和后代的产生来增殖基因型和转基因基因型。通过本发明的方法促进转基因植物或转基因猪胚胎，胎儿和后代的生产，因为供体核的分化细胞源可被遗传修饰和克隆繁殖。

9. 发明申请

US20060121607A1 Compositions and methods for neural differentiation of embryonic stem cells 审中-公开
标题翻译：胚胎干细胞神经分化的组成和方法
公开(公告)号：US20060121607A1
公开(公告)日：2006-06-08
申请号：US10524157
申请日：2003-08-08
申请人： Thomas Schulz , Steven Stice , Brian Condie , Bruce Davidson
发明人： Thomas Schulz , Steven Stice , Brian Condie , Bruce Davidson
IPC分类号： C12N5/08
CPC分类号： C12N5/0618 , A61K35/12 , C12N5/0619 , C12N2500/32 , C12N2500/90 , C12N2501/115 , C12N2501/91 , C12N2502/13 , C12N2506/02
摘要： The present invention provides compositions and methods for human neural cell production. More particularly, the present invention provides cellular differentiation methods employing an essentially serum free MEDII conditioned medium for the generation of human neural cells from pluripotent and multipotent human stem cells.
摘要翻译：本发明提供了人神经细胞产生的组合物和方法。更具体地，本发明提供了使用基本上无血清的MEDII条件培养基用于从多能和多能人干细胞产生人神经细胞的细胞分化方法。

10. 发明申请

US20060021070A1 Production of chimeric bovine or porcine animals using cultured inner cell mass 审中-公开
标题翻译：使用培养的内细胞块生产嵌合牛或猪动物
公开(公告)号：US20060021070A1
公开(公告)日：2006-01-26
申请号：US11024010
申请日：2004-12-28
申请人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. Leon , D. Jerry
发明人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. Leon , D. Jerry
IPC分类号： A01K67/027
CPC分类号： C12N15/8771 , A01K2217/05 , C12N15/8778
摘要： Novel cultured inner cell mass (CICM) cells, and cell lines, derived from ungulates, in particular, pigs and cows, and methods for their preparation are provided. The subject CICMs possess similar morphology and express cell markers identically or substantially similarly to ICMs of undifferentiated developing embryos for prolonged culturing periods. Heterologous DNA is inserted into the subject CICM cells and cell lines so as produce transgenic CICM cell which are introduced into non-human fertilized embryos to produce transgenic chimeric embryos. The transgenic chimeric embryos are transferred into recipient females where they are permitted to develop into transgenic chimeric fetuses. Recipient females give birth to transgenic chimeric animals which are capable of transmitting the heterologous DNA to their progeny. Transgenic CICM cells are also used to produce cloned transgenic embryos, fetuses and offspring.
摘要翻译：提供了新颖的培养的内细胞团（CICM）细胞和源自有蹄类动物，特别是猪和牛的细胞系及其制备方法。受试者CICM具有与未分化发育的胚胎的ICM相似或基本上类似于延长培养期的相似形态和表达细胞标记。将异源DNA插入受试者CICM细胞和细胞系中，以产生转入非人受精胚胎以产生转基因嵌合胚胎的转基因CICM细胞。将转基因嵌合胚胎转移到受体雌性中，在其中允许其转化为转基因嵌合胎儿。受体女性产生能够将异源DNA转移到其后代的转基因嵌合动物。转基因CICM细胞也用于产生克隆的转基因胚胎，胎儿和后代。

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