公开(公告)号：US08753884B2

公开(公告)日：2014-06-17

申请号：US12152779

申请日：2008-05-16

申请人： Jose Cibelli ,  Michael D. West ,  Robert Lanza

发明人： Jose Cibelli ,  Michael D. West ,  Robert Lanza

IPC分类号： C12N5/00 ,  C12N5/02 ,  C12N15/00

CPC分类号： C12N5/16 ,  A61K35/12 ,  C12N5/0606 ,  C12N2510/00

摘要： An improved method of producing differentiated progenitor cells comprising obtaining inner cell mass cells from a blastocyst and inducing differentiation of the inner cell mass cells to produce differentiated progenitor cells. The differentiated progenitor cells may be transfected such that there is an addition, deletion or alteration of a desired gene. The differentiated progenitor cells are useful in cell therapy and as a I source of cells for the production of tissues and organs for transplantation. Also provided is a method of producing a lineage-defective human embryonic stem cell.

摘要翻译： 一种改进的产生分化祖细胞的方法，包括从囊胚获得内细胞质量细胞并诱导内细胞大量细胞的分化以产生分化祖细胞。 可以转染分化的祖细胞，使得存在所需基因的添加，缺失或改变。 分化的祖细胞可用于细胞治疗和作为用于产生移植组织和器官的细胞的I来源。 还提供了产生谱系缺陷型人胚胎干细胞的方法。

公开(公告)号：US06808704B1

公开(公告)日：2004-10-26

申请号：US09655815

申请日：2000-09-06

申请人： Robert Lanza ,  Michael D. West ,  Jose Cibelli

发明人： Robert Lanza ,  Michael D. West ,  Jose Cibelli

IPC分类号： A01N6300

CPC分类号： C12N15/873 ,  A01K67/0271 ,  A01K2217/05 ,  A61K35/12 ,  A61K49/0008 ,  C12N15/8771 ,  C12N2517/04

摘要： This invention relates to methods for making immune compatible tissues and cells for the purpose of transplantation and tissue engineering, using the techniques of nuclear transfer and cloning. Also encompassed are methods for determining the effect on immune compatibility of expressed transgenes and other genetic manipulations of the engineered cells and tissues.

摘要翻译： 本发明涉及使用核转移和克隆技术制备用于移植和组织工程的免疫相容性组织和细胞的方法。 还包括确定表达的转基因对工程细胞和组织的免疫相容性和其他遗传操作的影响的方法。

公开(公告)号：US07696404B2

公开(公告)日：2010-04-13

申请号：US10329979

申请日：2002-12-27

申请人： Jose Cibelli ,  Michael D. West

发明人： Jose Cibelli ,  Michael D. West

IPC分类号： C12N15/00 ,  C12N5/02

CPC分类号： C12N15/873 ,  A01K2217/075 ,  A61K35/12 ,  A61K38/00 ,  C12N15/87 ,  C12N15/877 ,  C12N2510/00 ,  C12N2517/04

摘要： An improved method of nuclear transfer involving the transplantation of differentiated 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. Also, methods for improving nuclear transfer efficiency by genetically altering donor cells to inhibit apoptosis, select for a specific cell cycle and/or enhance embryonic growth and development are provided.

摘要翻译： 提供了涉及将不同供体细胞核移植到与供体细胞不同的物种的去核卵母细胞中的改进的核转移方法。 所得到的核转移单位可用于生产等原胚胎干细胞，特别是人原代胚胎干细胞。 这些胚胎或干细胞样细胞可用于产生所需的分化细胞并用于通过同源重组引入，去除或修饰所需基因，例如在这些细胞的基因组的特定位点。 这些可能含有异源基因的细胞特别适用于细胞移植治疗和体外研究细胞分化。 此外，提供了通过遗传改变供体细胞以抑制细胞凋亡，选择特定细胞周期和/或增强胚胎生长和发育来提高核转移效率的方法。

公开(公告)号：US20090104697A1

公开(公告)日：2009-04-23

申请号：US12152779

申请日：2008-05-16

申请人： Jose Cibelli ,  Michael D. West ,  Robert Lanza

发明人： Jose Cibelli ,  Michael D. West ,  Robert Lanza

IPC分类号： C12N5/02

CPC分类号： C12N5/16 ,  A61K35/12 ,  C12N5/0606 ,  C12N2510/00

摘要： An improved method of producing differentiated progenitor cells comprising obtaining inner cell mass cells from a blastocyst and inducing differentiation of the inner cell mass cells to produce differentiated progenitor cells. The differentiated progenitor cells may be transfected such that there is an addition, deletion or alteration of a desired gene. The differentiated progenitor cells are useful in cell therapy and as a I source of cells for the production of tissues and organs for transplantation. Also provided is a method of producing a lineage-defective human embryonic stem cell.

摘要翻译： 一种改进的产生分化祖细胞的方法，包括从囊胚获得内细胞质量细胞并诱导内细胞大量细胞的分化以产生分化祖细胞。 可以转染分化的祖细胞，使得存在所需基因的添加，缺失或改变。 分化的祖细胞可用于细胞治疗和作为用于产生移植组织和器官的细胞的I来源。 还提供了产生谱系缺陷型人胚胎干细胞的方法。

公开(公告)号：US20090136464A1

公开(公告)日：2009-05-28

申请号：US12218045

申请日：2008-07-09

申请人： Jose Cibelli ,  Tanja Dominko

发明人： Jose Cibelli ,  Tanja Dominko

IPC分类号： A61K35/12 ,  A61P43/00

CPC分类号： A01K67/0271 ,  A01K2217/05 ,  A61K35/12 ,  A61K48/00 ,  C12N15/873 ,  C12N2517/02 ,  C12N2517/04

摘要： Improved methods of cell therapy are provided using cells and tissues that are histocompatible with a human or non-human transplant recipient. The cells and tissues for transplant produced by the present invention exhibit a youthful state and can be committed to specific cell lineages to better infiltrate and proliferate at a desired target, e.g., a tissue, or organ in need of cell replacement therapy. For providing cells and tissues for transplant to a non-human mammal, the cells and tissues can be isolated from a gastrulating embryo produced by same-species nuclear transfer. Histocompatible cells and tissues for transplant to a human can be isolated from a gastrulating embryo that (i) is genetically modified to be in capable of developing beyond an early stage, or (ii) is produced by cross-species nuclear transfer between a human nuclear donor cell and an enucleated recipient cell, e.g., an oocyte, of a non-human mammal, or (iii) is produced by androgenesis or gynogenesis, or from pluripotent stem cells generated from such an embryo. Methods for producing histocompatible cells and tissues for transplant to a human can also be used to produce such cells or tissues for transplant to non-human mammals. The present invention also provides model embryonic systems having defined genetic makeup that are useful for developing and testing methods for cell and tissue therapy, and for studying genetic imprinting, reprogramming, rejuvenation, and other biochemical, metabolic, and physiological phenomena associated with embryogenesis.

摘要翻译： 使用与人或非人类移植受体组织相容的细胞和组织提供改进的细胞治疗方法。 本发明生产的用于移植的细胞和组织表现出年轻状态，并且可以致力于特定的细胞谱系，以在期望的靶，例如需要细胞替代疗法的组织或器官中更好地渗透和增殖。 为了提供用于移植到非人哺乳动物的细胞和组织，细胞和组织可以从通过相同种类的核转移产生的诱导胚胎中分离出来。 用于移植到人中的组织相容的细胞和组织可以从一个起胚胚胎中分离出来，（i）经过基因修饰以能够在早期阶段发育，或（ii）通过人类核之间的跨物种核转移产生 供体细胞和去核的受体细胞，例如非人哺乳动物的卵母细胞，或（iii）通过雄激素或雌激素产生，或由这种胚胎产生的多能干细胞产生。 用于生产用于移植到人中的组织相容性细胞和组织的方法也可以用于产生用于移植到非人哺乳动物的这种细胞或组织。 本发明还提供具有确定的遗传构成的模型胚胎系统，其可用于细胞和组织治疗的开发和测试方法，以及用于研究与胚胎发生相关的遗传印记，重编程，复壮和其他生物化学，代谢和生理现象。

公开(公告)号：US20060240556A1

公开(公告)日：2006-10-26

申请号：US11473729

申请日：2006-06-22

申请人： Jose Cibelli

发明人： Jose Cibelli

IPC分类号： C12N15/00

CPC分类号： C12N15/873 ,  A61K35/12 ,  C12N2517/04

摘要： Methods for making human ES cells and human differentiated cells and tissues for transplantation are described, whereby the cells and tissues are created following somatic cell nuclear transfer. The nuclear transfer donor is genetically modified prior to nuclear transfer such that cells of at least one developmental lineage are de-differentiated, i.e., unable to develop, thereby resolving the ethical dilemmas involved in reprogramming somatic cells back to the embryonic stage. The method concomitantly directs differentiation such that the desired cells and tissues may be more readily isolated.

公开(公告)号：US20060083722A1

公开(公告)日：2006-04-20

申请号：US10484398

申请日：2002-01-25

申请人： Jose Cibelli ,  Tanja Dominko

发明人： Jose Cibelli ,  Tanja Dominko

IPC分类号： A61K48/00 ,  C12N5/08

CPC分类号： A01K67/0271 ,  A01K2217/05 ,  A61K35/12 ,  A61K48/00 ,  C12N15/873 ,  C12N2517/02 ,  C12N2517/04

摘要： Improved methods of cell therapy are provided using cells and tissues that are histocompatible with a human or non-human mammal transplant recipient. The cells and tissues for transplant produced by the present invention exhibit a youthful state and can be committed to specific cell lineages to better infiltrate and proliferate at a desired target, e.g., a tissue, or organ in need of cell replacement therapy. For providing cells and tissues for transplant to a non-human mammal, the cells and tissues can be isolated from a gastrulating embryo produced by same-species nuclear transfer. Histocompatible cells and tissues for transplant to a human can be isolated from a gastrulating embryo that (i) is genetically modified to be in capable of developing beyond and early stage, or (ii) is produced by cross-species nuclear transfer between a human nuclear donor cell and an enucleated recipient cell, e.g., an oocyte, of a non-human mammal, or (iii) is produced by androgenesis or gynogenesis, or from pluripotent stem cells generated from such an embryo. Methods for producing histocompatible cells and tissues for transplant to a human can also be used to produce such cells or tissues for transplant to non-human mammals. The present invention also provides model embryonic systems having defined genetic makeup that are useful for developing and testing methods for cell and tissue therapy, and for studying genetic imprinting, reprogramming, rejuvenation, and other biochemical, metabolic, and physiological phenomena associated with embryogenesis.

摘要翻译： 使用与人或非人哺乳动物移植受体组织相容的细胞和组织提供改进的细胞治疗方法。 本发明生产的用于移植的细胞和组织表现出年轻状态，并且可以致力于特定的细胞谱系，以在期望的靶，例如需要细胞替代疗法的组织或器官中更好地渗透和增殖。 为了提供用于移植到非人哺乳动物的细胞和组织，细胞和组织可以从通过相同种类的核转移产生的诱导胚胎中分离出来。 用于移植到人的组织相容的细胞和组织可以从一个起胚胚胎中分离出来，其中（i）经遗传修饰以能够发展到超前和早期阶段，或者（ii）是通过人类核之间的跨物种核转移产生的 供体细胞和去核的受体细胞，例如非人哺乳动物的卵母细胞，或（iii）通过雄激素或雌激素产生，或由这种胚胎产生的多能干细胞产生。 用于生产用于移植到人中的组织相容性细胞和组织的方法也可以用于产生用于移植到非人哺乳动物的这种细胞或组织。 本发明还提供具有确定的遗传构成的模型胚胎系统，其可用于细胞和组织治疗的开发和测试方法，以及用于研究与胚胎发生相关的遗传印记，重编程，复壮和其他生物化学，代谢和生理现象。

公开(公告)号：US06235970B1

公开(公告)日：2001-05-22

申请号：US08935052

申请日：1997-09-22

申请人： Steven L. Stice ,  Jose Cibelli ,  James Robl ,  Paul Golueke ,  F. Abel Ponce de Leon ,  D. Joseph Jerry

发明人： Steven L. Stice ,  Jose Cibelli ,  James Robl ,  Paul Golueke ,  F. Abel Ponce de Leon ,  D. Joseph Jerry

IPC分类号： C12N1500

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。

公开(公告)号：US20110244464A1

公开(公告)日：2011-10-06

申请号：US12974781

申请日：2010-12-21

申请人： Jose Cibelli ,  Javier Crosby ,  Emilio Fernandez ,  Arif Kocabas ,  Guilherme Jordao De Rosa

发明人： Jose Cibelli ,  Javier Crosby ,  Emilio Fernandez ,  Arif Kocabas ,  Guilherme Jordao De Rosa

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6883 ,  C12Q2600/158

摘要： A genetic means of determining whether a female subject produces “pregnancy competent” oocytes is provided. The means comprises detecting the level of expression of one or more genes that are expressed at characteristic levels (upregulated or downregulated) in cumulus cells derived from pregnancy competent oocytes. This characteristic gene expression level, or pattern referred to herein as the “pregnancy signature”, also can be used to identify subjects with underlying conditions that impair or prevent the development of a viable pregnancy, e.g., pre-menopausal condition, other hormonal dysfunction, ovarian dysfunction, ovarian cyst, cancer or other cell proliferation disorder, autoimmune disease and the like. Microarrays containing “pregnancy signature” genes or corresponding polypeptides provide another preferred aspect of the invention. Still further, the subject invention can be used to derive animal models, e.g., non-human primate animal models, for the evaluation of the efficacy of putative female fertility treatments.

摘要翻译： 提供确定女性受试者是否产生“怀孕受精”卵母细胞的遗传手段。 该方法包括检测在源于妊娠合格卵母细胞的卵丘细胞中以特征水平（上调或下调）表达的一种或多种基因的表达水平。 这种特征性基因表达水平或本文称为“怀孕签名”的模式也可以用于鉴定有潜在病症的受试者，其损害或预防可行妊娠的发展，例如绝经前期，其他激素功能障碍， 卵巢功能障碍，卵巢囊肿，癌症或其他细胞增殖障碍，自身免疫性疾病等。 包含“怀孕特征”基因或相应多肽的微阵列提供本发明的另一优选方面。 此外，本发明可以用于衍生动物模型，例如非人灵长类动物模型，以评估推定的女性生育力治疗的功效。

公开(公告)号：US20070054289A1

公开(公告)日：2007-03-08

申请号：US11437797

申请日：2006-05-22

申请人： Jose Cibelli ,  Javier Crosby ,  Emilio Fernandez ,  Arif Kocabas ,  Guilherme Jordao De Magalhaes Rosa

发明人： Jose Cibelli ,  Javier Crosby ,  Emilio Fernandez ,  Arif Kocabas ,  Guilherme Jordao De Magalhaes Rosa

IPC分类号： C12Q1/00 ,  C12Q1/68

CPC分类号： C12Q1/6883 ,  C12Q1/6881 ,  C12Q2600/158

摘要： A genetic means of determining whether a female subject produces “pregnancy competent” oocytes is provided. The means comprises detecting the level of expression of one or more genes that are expressed at characteristic levels (upregulated or downregulated) in cumulus cells derived from pregnancy competent oocytes. This characteristic gene expression level, or pattern referred to herein as the “pregnancy signature”, also can be used to identify subjects with underlying conditions that impair or prevent the development of a viable pregnancy, e.g., pre-menopausal condition, other hormonal dysfunction, ovarian dysfunction, ovarian cyst, cancer or other cell proliferation disorder, autoimmune disease and the like. Microarrays containing “pregnancy signature” genes or corresponding polypeptides provide another preferred aspect of the invention. Still further, the subject invention can be used to derive animal models, e.g., non-human primate animal models, for the evaluation of the efficacy of putative female fertility treatments. Additionally, an improved RNA amplification protocol is provided herein referred to as the CRL amplification protocol which is suitable for reproducibly amplifying all the RNAs expressed by a cell sample, even when only a few cells are available.

摘要翻译： 提供确定女性受试者是否产生“怀孕受精”卵母细胞的遗传手段。 该方法包括检测在源于妊娠合格卵母细胞的卵丘细胞中以特征水平（上调或下调）表达的一种或多种基因的表达水平。 这种特征性基因表达水平或本文称为“怀孕签名”的模式也可以用于鉴定有潜在病症的受试者，其损害或预防可行妊娠的发展，例如绝经前期，其他激素功能障碍， 卵巢功能障碍，卵巢囊肿，癌症或其他细胞增殖障碍，自身免疫性疾病等。 包含“怀孕特征”基因或相应多肽的微阵列提供本发明的另一优选方面。 此外，本发明可以用于衍生动物模型，例如非人灵长类动物模型，以评估推定的女性生育力治疗的功效。 此外，本文提供了改进的RNA扩增方案，其被称为CRL扩增方案，其适用于可再现地扩增由细胞样品表达的所有RNA，即使只有少数细胞可用。