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1. 发明申请

WO0043498A2 HUMAN LIVER PROGENITORS 审中-公开
标题翻译：人类肝脏发育者
公开(公告)号：WO0043498A2
公开(公告)日：2000-07-27
申请号：PCT/US0001116
申请日：2000-01-19
申请人： UNIV NORTH CAROLINA
发明人： REID LOLA M , KUBOTA HIROSHI , MOSS NICHOLAS
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
CPC分类号： C12N5/0672 , A61K35/12 , 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 mumber 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 expess 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 6-15 mu in diameter, diploid, glycophorin A , CD45 , AFP , ALB , ICAM and with subpopulations varying in expression of CD 14 , CD34 , CD38 , CD117 . These progenitor subpopulations have characteristics expected for cells that are particularly useful in liver cell and gene therapies and for extablishing bioartificial organs.
摘要翻译：公开了从人肝中分离和冷冻保存祖细胞的方法，其包括处理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; 使悬浮液进行减量步骤，其基本上减少悬液中非祖细胞的数量，并且提供富含祖细胞的减灭悬浮液，其表现出与一种或多种细胞谱系中的至少一种相关的一种或多种标记; 并且从所述切除的悬浮液中选择那些本身，其后代或其更成熟形式的那些细胞排出一种或多种与所述一种或多种细胞谱系中的至少一种相关的标记。这些标记物中有CD14，CD34，CD38，CD45和ICAM。肝祖细胞的特征为直径为6-15μm，二倍体，血型糖蛋白A - ，CD45 - ，AFP +++，ALB +，ICAM +和具有不同CD表达的亚群 14 +，CD34 ++，CD38 ++，CD117 +。这些祖细胞亚群具有在肝细胞和基因疗法中特别有用并且用于建立生物人造器官的细胞的预期特征。

2. 发明申请

WO2007140243A2 HEPATIC STELLATE CELL PRECURSORS AND METHODS OF ISOLATING SAME 审中-公开
标题翻译： HEPATIC STELLATE CELL PRECURSORS和隔离方法
公开(公告)号：WO2007140243A2
公开(公告)日：2007-12-06
申请号：PCT/US2007069645
申请日：2007-05-24
申请人： UNIV NORTH CAROLINA , KUBOTA HIROSHI , REID LOLA M
发明人： KUBOTA HIROSHI , REID LOLA M
IPC分类号： C12N5/00 , C12N5/071 , C12N5/074
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-I+, VCAM-I+, ß3-integrin+. In addition to expression of these surface markers, the cells also express the intracellular markers desmin, vimentin, smooth muscle a-actin, nestin, hepatocyte growth factor, stromal derived factor- la and Hlx 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-1a和Hlx同源异体转录因子。

3. 发明申请

WO0228997A3 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：WO0228997A3
公开(公告)日：2002-09-12
申请号：PCT/US0027429
申请日：2000-10-03
申请人： UNIV NORTH CAROLINA
发明人： KUBOTA HIROSHI , REID LOLA M
IPC分类号： G01N33/48 , A61K35/12 , A61K35/407 , A61K38/00 , A61K48/00 , A61P1/16 , C12N5/074 , G01N33/483 , G01N33/53 , C12N5/06 , C12N5/08 , C12N5/10
CPC分类号： C12N5/0672 , A61K35/12 , A61K48/00 , C12N2501/11 , C12N2502/13
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粒细胞或细胞质液滴的度量的侧向散射水平，其高于非实质细胞例如造血细胞，并且低于成熟实质细胞如肝细胞中。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。

4. 发明申请

WO0043498A9 HUMAN LIVER PROGENITORS 审中-公开
标题翻译：人类生殖器
公开(公告)号：WO0043498A9
公开(公告)日：2001-10-18
申请号：PCT/US0001116
申请日：2000-01-19
申请人： UNIV NORTH CAROLINA
发明人： REID LOLA M , KUBOTA HIROSHI , MOSS NICHOLAS
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 , A01N1/02 , C12N5/10
CPC分类号： C12N5/0672 , A61K35/12 , 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 mumber 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 expess 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 6-15 mu in diameter, diploid, glycophorin A , CD45 , AFP , ALB , ICAM and with subpopulations varying in expression of CD 14 , CD34 , CD38 , CD117 . These progenitor subpopulations have characteristics expected for cells that are particularly useful in liver cell and gene therapies and for extablishing 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数量的非祖细胞，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谱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，它们的子代或更成熟的形式表现与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。肝祖细胞的特征是直径为6-15μm，二倍体，血型糖蛋白A，CD45，AFP <+++>，ALB +，ICAM +和亚群在CD 14 +，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生物人造器官的细胞具有预期的特征。

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