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1. 发明申请

US20050148072A1 Human liver progenitors 审中-公开
标题翻译：人肝脏祖细胞
公开(公告)号：US20050148072A1
公开(公告)日：2005-07-07
申请号：US10944919
申请日：2004-09-21
申请人： Lola Reid , Nicholas Moss , Hiroshi Kubota
发明人： Lola Reid , Nicholas Moss , Hiroshi Kubota
IPC分类号： G01N33/53 , A61K35/12 , A61K35/407 , A61K48/00 , A61P1/16 , A61P7/00 , A61P35/00 , C12N5/00 , C12N5/074 , C12N15/09 , C12Q1/02 , C12Q1/68 , C12N5/08
CPC分类号： C12N5/0672 , A61K35/12 , A61K2039/55594 , C12N2500/20 , C12N2500/22 , C12N2500/25 , C12N2500/36 , C12N2503/02
摘要： Methods of isolating and cryopreserving progenitors from human liver are disclosed which include processing human liver tissue to provide a substantially single cell suspension comprising progenitors and non-progenitors of one or more cell lineages found in human liver; subjecting the suspension to a debulking step, which reduces substantially the number of non-progenitors in the suspension, and which provides a debulked suspension enriched in progenitors exhibiting one or more markers associated with at least one of the one or more cell lineages; and selecting from said debulked suspension those cells, which themselves, their progeny, or more mature forms thereof express one or more markers associated with at least one of the one or more cell lineages. Among these markers are CD14, CD34, CD38, CD45, and ICAM. Hepatic progenitors are characterized as being 6-15μ in diameter, diploid, glycophorin A−, CD45−, AFP+++, ALB+, ICAM+, and with subpopulations varying in expression of CD14+. CD34++, CD38++, CD117+. These progenitor subpopulations have characteristics expected for cells that are particularly useful in liver cell and gene therapies and for establishing bioartificial organs.
摘要翻译：公开了从人肝脏分离和冷冻保存祖细胞的方法，其包括加工人肝组织以提供包含在人肝脏中发现的一种或多种细胞谱系的祖细胞和非祖细胞的基本上单细胞悬浮液; 使悬浮液经历减压步骤，其减少了悬浮液中的非祖细胞的数量，并且提供了富含前体细胞的减数分裂的悬浮液，其显示出与一种或多种细胞系中的至少一种相关联的一种或多种标志物; 以及从所述减粘悬浮液中选择这些细胞，其本身，其子代或更成熟的形式表达与所述一种或多种细胞谱系中的至少一种相关联的一种或多种标志物。这些标记物中有CD14，CD34，CD38，CD45和ICAM。肝祖细胞的特征在于直径为6-15μm，二倍体，血型糖蛋白，CD45 - ，AFP +++，ALB + > + ，ICAM ，以及与CD14 + + / / SUP的表达变化的亚群。 CD34 ++ ++，CD38 ++，CD117 + + / / 这些祖细胞亚群对于在肝细胞和基因治疗中特别有用的细胞以及用于建立生物人造器官的细胞具有预期的特征。

2. 发明申请

US20070238172A1 METHODS OF ISOLATING BIPOTENT HEPATIC PROGENITOR CELLS 审中-公开
标题翻译：分离BIPOTEHHEPATIC PROGENITOR细胞的方法
公开(公告)号：US20070238172A1
公开(公告)日：2007-10-11
申请号：US11757168
申请日：2007-06-01
申请人： Hiroshi Kubota , Lola Reid
发明人： Hiroshi Kubota , Lola Reid
IPC分类号： C12N5/06
CPC分类号： G01N33/5091 , C12N5/0672 , C12N2501/50 , C12N2501/58 , C12N2501/599
摘要： A method of obtaining a mixture of cells enriched in hepatic progenitors is developed which comprises methods yielding suspensions of a mixture of cell types, and selecting those cells that are classical MHC class I antigen(s) negative and ICAM-1 antigen positive. The weak or dull expression of nonclassical MHC class I antigen(s) can be used for further enrichment of hepatic progenitors. Furthermore, the progenitors can be selected to have a level of side scatter, a measure of granularity or cytoplasmic droplets, that is higher than that in non-parenchymal cells, such as hemopoietic cells, and lower than that in mature parenchymal cells, such as hepatocytes. Furthermore, the progeny of the isolated progenitors can express alpha-fetoprotein and/or albumin and/or CK19. The hepatic progenitors, so isolated, can grow clonally, that is an entire population of progeny can be derived from one cell. The clones of progenitors have a growth pattern in culture of piled-up aggregates or clusters. These methods of isolating the hepatic progenitors are applicable to any vertebrates including human. The hepatic progenitor cell population is expected to be useful for cell therapies, for bioartificial livers, for gene therapies, for vaccine development, and for myriad toxicological, pharmacological, and pharmaceutical programs and investigations.
摘要翻译：开发了获得富含肝祖细胞的细胞混合物的方法，其包括产生细胞类型混合物的悬浮液的方法，并选择那些经典的MHC I类抗原和ICAM-1抗原阳性的细胞。非典型MHC I类抗原的弱或暗的表达可用于进一步富集肝祖细胞。此外，可以选择祖细胞具有高于非实质细胞例如造血细胞的侧向散射，粒度或细胞质液滴的量级，并且低于成熟实质细胞中的祖细胞，例如肝细胞。此外，分离的祖细胞的后代可以表达甲胎蛋白和/或白蛋白和/或CK19。如此分离的肝祖细胞可以克隆生长，即可以从一个细胞中获得后代的全部群体。祖先的克隆在堆积聚集体或簇的培养中具有增长模式。这些分离肝祖细胞的方法适用于包括人在内的任何脊椎动物。预期肝祖细胞群可用于细胞疗法，生物人工肝，基因治疗，疫苗开发和无数毒理学，药理学和药物程序和调查。

3. 发明申请

US20080131900A1 VARIANTS OF ALPHA-FETOPROTEIN CODING AND EXPRESSION SEQUENCES 有权
标题翻译： ALPHA-FETOPROTEIN编码和表达序列的变体
公开(公告)号：US20080131900A1
公开(公告)日：2008-06-05
申请号：US11963207
申请日：2007-12-21
申请人： Hiroshi Kubota , Robert Storms , Lola Reid
发明人： Hiroshi Kubota , Robert Storms , Lola Reid
IPC分类号： C12Q1/68 , C07K16/00 , C12N5/00
CPC分类号： C07K14/4715
摘要： The invention discloses the sequences of variant forms of alpha-fetoprotein transcripts that have been identified in human hemopoietic progenitors but not in differentiated mature cells. The variant forms of AFP (vAFP) cDNA sequences isolated from a multipotent hemopoietic cell line, K562, differ from the authentic AFP transcript, consisting of 15 exons, by lacking only exon 1. Instead of exon 1, vAFP transcripts use an additional one or two exons located in the 5′-untranslated region of the AFP gene. K562 expressed selectively vAFP, whereas a hepatocellular carcinoma cell line, HepG2, showed no detectable expression of vAFP. In normal adult tissues, vAFP transcripts is detected in the bone marrow, thymus and brain, but not the spleen, suggesting the expression occurs in normal hemopoietic progenitors. Moreover, CD34+Lin− hemopoietic stem/progenitor cells purified by flow cytometric sorting also express the variant transcripts.
摘要翻译：本发明公开了已经在人造血祖细胞中鉴定但不在分化的成熟细胞中的甲胎蛋白转录物的变体形式的序列。从多能造血细胞系K562分离的AFP（vAFP）cDNA序列的变体形式与仅由外显子1缺失的15个外显子组成的真正的AFP转录物不同。代替外显子1，vAFP转录物使用另外一个或位于AFP基因的5'非翻译区的两个外显子。 K562选择性表达vAFP，而肝细胞癌细胞系HepG2则显示无可检测的vAFP表达。在正常的成年组织中，在骨髓，胸腺和脑中检测到vAFP转录物，而不是脾脏，表明在正常造血祖细胞中发生表达。此外，通过流式细胞分选纯化的CD34 +造血干细胞/祖细胞也表达变体转录物。

4. 发明申请

US20080009057A1 HEPATIC STELLATE CELL PRECURSORS AND METHODS OF ISOLATING SAME 有权
标题翻译： HEPATIC STELLATE CELL PRECURSORS和隔离方法
公开(公告)号：US20080009057A1
公开(公告)日：2008-01-10
申请号：US11753326
申请日：2007-05-24
申请人： Hiroshi Kubota , Lola Reid
发明人： Hiroshi Kubota , Lola Reid
IPC分类号： C12N5/00
CPC分类号： C12N5/0018 , C12N5/067 , C12N5/0672 , C12N2500/00 , C12N2500/05 , C12N2500/20 , C12N2500/25 , C12N2500/32 , C12N2500/36 , C12N2500/38 , C12N2500/90 , C12N2501/00 , C12N2501/11 , C12N2501/235 , C12N2501/33 , G01N33/56966
摘要： The present invention relates to precursor cells to hepatic stellate cells, compositions comprising same and methods of isolating same. The surface antigenic profile of the precursors is MHC class Ia negative, ICAM-1+, VCAM-1+, β3-integrin+. In addition to expression of these surface markers, the cells also express the intracellular markers desmin, vimentin, smooth muscle α-actin, nestin, hepatocyte growth factor, stromal derived factor-1α and H1x homeobox transcriptional factor.
摘要翻译：本发明涉及肝星状细胞的前体细胞，包含其的组合物及其分离方法。前体的表面抗原谱是MHC Ia类阴性，ICAM-1 +，VCAM-1 +，β3-整联蛋白+。除了这些表面标志物的表达外，细胞还表达细胞内标志物结蛋白，波形蛋白，平滑肌α-肌动蛋白，巢蛋白，肝细胞生长因子，基质衍生因子-1α和H1x同源异体转录因子。

5. 发明申请

US20070231853A1 METHODS OF ISOLATING BIPOTENT HEPATIC PROGENITOR CELLS 审中-公开
公开(公告)号：US20070231853A1
公开(公告)日：2007-10-04
申请号：US11758593
申请日：2007-06-05
申请人： Hiroshi Kubota , Lola Reid
发明人： Hiroshi Kubota , Lola Reid
IPC分类号： C12Q1/06 , C12N5/08
CPC分类号： G01N33/5091 , C12N5/0672 , C12N2501/50 , C12N2501/58 , C12N2501/599
摘要： A method of obtaining a mixture of cells enriched in hepatic progenitors is developed which comprises methods yielding suspensions of a mixture of cell types, and selecting those cells that are classical MHC class I antigen(s) negative and ICAM-1 antigen positive. The weak or dull expression of nonclassical MHC class I antigen(s) can be used for further enrichment of hepatic progenitors. Furthermore, the progenitors can be selected to have a level of side scatter, a measure of granularity or cytoplasmic droplets, that is higher than that in non-parenchymal cells, such as hemopoietic cells, and lower than that in mature parenchymal cells, such as hepatocytes. Furthermore, the progeny of the isolated progenitors can express alpha-fetoprotein and/or albumin and/or CK19. The hepatic progenitors, so isolated, can grow clonally, that is an entire population of progeny can be derived from one cell. The clones of progenitors have a growth pattern in culture of piled-up aggregates or clusters. These methods of isolating the hepatic progenitors are applicable to any vertebrates including human. The hepatic progenitor cell population is expected to be useful for cell therapies, for bioartificial livers, for gene therapies, for vaccine development, and for myriad toxicological, pharmacological, and pharmaceutical programs and investigations.

6. 发明申请

US20070042341A1 Biodegradable polymer-ligand conjugates and their uses in isolation of cellular subpopulations and in cryopreservation, culture and transplantation of cells 审中-公开
标题翻译：生物可降解聚合物 - 配体结合物及其在细胞亚群分离中的用途和细胞的低温保存，培养和移植
公开(公告)号：US20070042341A1
公开(公告)日：2007-02-22
申请号：US11484615
申请日：2006-07-12
申请人： Arron Xu , Lola Reid
发明人： Arron Xu , Lola Reid
IPC分类号： A01N1/02 , G01N33/567 , C12N5/08
CPC分类号： C12N5/0672 , C12N2533/30 , C12N2533/40 , C12N2533/54
摘要： The invention discloses a biodegradable particle-cell composition having at least one biodegradable particle, at least one receptive group covalently linked thereto, and a cell anchored thereto. The particle can be polylactide, a polylactide-lysine copolymer, polylactide-lysine-polyethylene glycol copolymer, starch, or collagen. The receptive group can be an antibody, a fragment of an antibody, an avidin, a streptavidin, or a biotin moiety. Moreover, the particle can also have extracellular matrix components other than collagen. The particle-cell compositions can be used for selection of cells from a population, for cell culture of anchorage-dependent cells, for cryopreservation of anchorage-dependent cells, and for transplantation as a cell therapy.
摘要翻译：本发明公开了一种具有至少一种可生物降解的颗粒，至少一种与其共价连接的接受基团的可生物降解的颗粒 - 细胞组合物和锚定于其上的细胞。该颗粒可以是聚丙交酯，聚丙交酯 - 赖氨酸共聚物，聚丙交酯 - 赖氨酸 - 聚乙二醇共聚物，淀粉或胶原蛋白。接受组可以是抗体，抗体的片段，抗生物素蛋白，链霉亲和素或生物素部分。此外，颗粒还可以具有除胶原之外的细胞外基质成分。颗粒细胞组合物可用于选择来自群体的细胞，用于锚定依赖性细胞的细胞培养，用于冷冻保存锚定依赖性细胞的细胞培养，以及用于细胞治疗的移植。

7. 发明申请

US20070155009A1 EXTRACELLULAR MATRIX COMPONENTS FOR EXPANSION OR DIFFERENTIATION OF HEPATIC PROGENITORS 审中-公开
标题翻译：用于扩增或分化肝脏原发性细胞的细胞基质组分
公开(公告)号：US20070155009A1
公开(公告)日：2007-07-05
申请号：US11560049
申请日：2006-11-15
申请人： Randall McClelland , Lola Reid
发明人： Randall McClelland , Lola Reid
IPC分类号： C12N5/06 , C12N5/08 , C12M3/00
CPC分类号： C12N5/0672 , C12N2500/90 , C12N2502/02 , C12N2502/03 , C12N2502/14 , C12N2533/50 , C12N2533/52 , C12N2533/54 , C12N2533/70 , C12N2533/90
摘要： A method is provided of propagating hepatic progenitors in vitro on or in one or multiple extracellular matrix components found in the stem cell compartment or niche of liver. A container for the propagation of the progenitors and comprising culture dishes, bioreactors, or lab chips.
摘要翻译：提供了一种在肝脏的干细胞隔室或利基中发现的一种或多种细胞外基质组分上或在体外传播肝祖细胞的方法。用于繁殖祖细胞并包含培养皿，生物反应器或实验室碎片的容器。

8. 发明申请

US20070099297A1 PRIMITIVE AND PROXIMAL HEPATIC STEM CELLS 有权
标题翻译：主要和临床干细胞
公开(公告)号：US20070099297A1
公开(公告)日：2007-05-03
申请号：US11609729
申请日：2006-12-12
申请人： Lola Reid , Nick Moss , Mark Furth , John Ludlow , Andrew Bruce
发明人： Lola Reid , Nick Moss , Mark Furth , John Ludlow , Andrew Bruce
IPC分类号： C12N5/08
CPC分类号： C12N5/0672 , C12N2500/25 , C12N2500/36 , C12N2500/90 , C12N2501/11 , C12N2502/13 , G01N33/5005 , G01N33/68 , G01N2333/705 , G01N2333/70525 , G01N2333/70596
摘要： Hepatic progenitors comprise two populations of human hepatic stem cells, primitive and proximal hepatic stem cells, and two populations of committed progenitors, one for biliary cells and one for hepatocytes. Human primitive hepatic stem cells are a very small fraction of the liver cell population and give rise to proximal hepatic stem cells constituting a much larger fraction of the liver. Human proximal hepatic stem cells give rise to biliary and hepatocyte committed progenitors. Primitive and proximal stem cells are the primary stem cells for the human liver. Human primitive hepatic stem cells may be isolated by immunoselection from human livers or culturing human liver cells under conditions which select for a human primitive hepatic stem cell. Proximal hepatic stem cells may be isolated by immunoselection, or by culturing human liver cells under conditions which include a developmental factor. Proximal hepatic stem cells may also be isolated by culturing colonies comprising a primitive hepatic stem cell under conditions which include a developmental factor. Resulting compositions may be used for treating liver disorders and for producing bioartificial organs.
摘要翻译：肝祖细胞包含两个人类肝干细胞，原始和近端肝干细胞，以及两个定型祖细胞群，一个为胆汁细胞，一个为肝细胞。人原始肝干细胞是肝细胞群体的非常小的一部分，并且引起近端肝干细胞构成肝脏的大部分。人近端肝干细胞产生胆汁和肝细胞定向的祖细胞。原始和近端干细胞是人类肝脏的主要干细胞。人原始肝干细胞可以通过人肝脏的免疫选择或在选择人原始肝干细胞的条件下培养人肝细胞来分离。可以通过免疫选择或通过在包括发育因子的条件下培养人肝细胞来分离近端肝干细胞。也可以通过在包括发育因子的条件下培养包含原始肝干细胞的菌落来分离近端肝干细胞。所得组合物可用于治疗肝脏疾病和用于生产生物人造器官。

9. 发明申请

US20050100877A1 Biodegradable polymer-ligand conjugates and their uses in isolation of cellular subpolulations and in cryopreservation, culture and transplantation of cells 审中-公开
标题翻译：生物可降解聚合物 - 配体共轭物及其在细胞分泌分离中的用途以及细胞的低温保存，培养和移植
公开(公告)号：US20050100877A1
公开(公告)日：2005-05-12
申请号：US10931073
申请日：2004-09-01
申请人： Arron Xu , Lola Reid
发明人： Arron Xu , Lola Reid
IPC分类号： A01N1/02 , A61K39/385 , A61K39/395 , A61K39/44 , C12M1/00 , C12N20060101 , C12N5/074 , C12N11/00 , C12N11/02
CPC分类号： C12N5/0672 , C12N2533/30 , C12N2533/40 , C12N2533/54
摘要： The invention discloses a biodegradable particle-cell composition having at least one biodegradable particle, at least one receptive group covalently linked thereto, and a cell anchored thereto. The particle can be polylactide, a polylactide-lysine copolymer, polylactide-lysine-polyethylene glycol copolymer, starch, or collagen. The receptive group can be an antibody, a fragment of an antibody, an avidin, a streptavidin, or a biotin moiety. Moreover, the particle can also have extracellular matrix components other than collagen. The particle-cell compositions can be used for selection of cells from a population, for cell culture of anchorage-dependent cells, for cryopreservation of anchorage-dependent cells, and for transplantation as a cell therapy.
摘要翻译：本发明公开了一种具有至少一种可生物降解的颗粒，至少一种与其共价连接的接受基团的可生物降解的颗粒 - 细胞组合物和锚定于其上的细胞。该颗粒可以是聚丙交酯，聚丙交酯 - 赖氨酸共聚物，聚丙交酯 - 赖氨酸 - 聚乙二醇共聚物，淀粉或胶原蛋白。接受组可以是抗体，抗体的片段，抗生物素蛋白，链霉亲和素或生物素部分。此外，颗粒还可以具有除胶原之外的细胞外基质成分。颗粒细胞组合物可用于选择来自群体的细胞，用于锚定依赖性细胞的细胞培养，用于冷冻保存锚定依赖性细胞的细胞培养，以及用于细胞治疗的移植。

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