METHODS AND COMPOSITIONS FOR DETECTING RARE CELLS FROM A BIOLOGICAL SAMPLE
    3.
    发明申请
    METHODS AND COMPOSITIONS FOR DETECTING RARE CELLS FROM A BIOLOGICAL SAMPLE 有权
    用于从生物样品中检测稀有细胞的方法和组合物

    公开(公告)号:US20080206757A1

    公开(公告)日:2008-08-28

    申请号:US11841972

    申请日:2007-08-20

    IPC分类号: C12Q1/58 C12N1/20 G01N33/53

    摘要: The present invention provides methods and compositions for isolating and detecting rare cells from a biological sample containing other types of cells. In particular, the present invention includes a debulking step that uses a microfabricated filters for filtering fluid samples and the enriched rare cells can be used in a downstream process such as identifies, characterizes or even grown in culture or used in other ways. The invention also include a method of determining the aggressiveness of the tumor or of the number or proportion of cancer cells in the enriched sample by detecting the presence or amount of telomerase activity or telomerase nucleic acid or telomerase expression after enrichment of rare cells. This invention further provides an efficient and rapid method to specifically remove red blood cells as well as white blood cells from a biological sample containing at least one of each of red blood cells and white blood cells, resulting in the enrichment of rare target cells including circulating tumor cells (CTC), stromal cells, mesenchymal cells, endothelial cells, fetal cells, stem cells, non-hematopoietic cells etc from a blood sample. The method is based upon combination of immuno-microparticles (antibody coated microparticles) and density-based separation. The final enriched target cells can be subjected to a variety of analysis and manipulations, such as flowcytometry, PCR, immunofluorescence, immunocytochemistry, image analysis, enzymatic assays, gene expression profiling analysis, efficacy tests of therapeutics, culturing of enriched rare cells, and therapeutic use of enriched rare cells. In addition, depleted plasma protein and white blood cells can be optionally recovered, and subjected to other analysis such as inflammation studies, gene expression profiling, etc.

    摘要翻译: 本发明提供从含有其他类型细胞的生物样品中分离和检测稀有细胞的方法和组合物。 特别地,本发明包括减压步骤,其使用微制造过滤器来过滤流体样品,并且富集的稀有细胞可以用于下游过程,例如识别,表征甚至在培养中生长或以其它方式使用。 本发明还包括通过在稀释细胞富集后检测端粒酶活性或端粒酶核酸或端粒酶表达的存在或数量来确定肿瘤侵袭性或富集样品中癌细胞数量或比例的方法。 本发明还提供了一种从含有红细胞和白细胞中的至少一种的生物样品中特异性除去红细胞以及白细胞的有效快速方法,导致稀有靶细胞的富集,包括循环 来自血液样品的肿瘤细胞(CTC),基质细胞,间充质细胞,内皮细胞,胎儿细胞,干细胞,非造血细胞等。 该方法基于免疫微粒(抗体包被的微粒)和基于密度的分离的组合。 最终的富集的靶细胞可以进行各种分析和操作,例如流式细胞术,PCR,免疫荧光,免疫细胞化学,图像分析,酶测定,基因表达谱分析,治疗剂的功效测试,培养富集的稀有细胞和治疗 使用富集的稀有细胞。 此外,可以任选地回收贫化的血浆蛋白和白细胞,并进行其他分析,例如炎症研究,基因表达谱等。

    METHODS AND COMPOSITIONS FOR DETECTING RARE CELLS FROM A BIOLOGICAL SAMPLE

    公开(公告)号:US20080057505A1

    公开(公告)日:2008-03-06

    申请号:US11777962

    申请日:2007-07-13

    摘要: The present invention provides methods and compositions for isolating and detecting rare cells from a biological sample containing other types of cells. In particular, the present invention includes a debulking step that uses a microfabricated filters for filtering fluid samples and the enriched rare cells can be used in a downstream process such as identifies, characterizes or even grown in culture or used in other ways. The invention also include a method of determining the aggressiveness of the tumor or of the number or proportion of cancer cells in the enriched sample by detecting the presence or amount of telomerase activity or telomerase nucleic acid or telomerase expression after enrichment of rare cells. This invention further provides an efficient and rapid method to specifically remove red blood cells as well as white blood cells from a biological sample containing at least one of each of red blood cells and white blood cells, resulting in the enrichment of rare target cells including circulating tumor cells (CTC), stromal cells, mesenchymal cells, endothelial cells, fetal cells, stem cells, non-hematopoietic cells etc from a blood sample. The method is based upon combination of immuno-microparticles (antibody coated microparticles) and density-based separation. The final enriched target cells can be subjected to a variety of analysis and manipulations, such as flowcytometry, PCR, immunofluorescence, immunocytochemistry, image analysis, enzymatic assays, gene expression profiling analysis, efficacy tests of therapeutics, culturing of enriched rare cells, and therapeutic use of enriched rare cells. In addition, depleted plasma protein and white blood cells can be optionally recovered, and subjected to other analysis such as inflammation studies, gene expression profiling, etc.