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1. 发明申请

WO2002062950A2 METHOD FOR QUANTIFICATION OF RECOMBINANT VIRUSES 审中-公开
标题翻译：重组病毒的定量方法
公开(公告)号：WO2002062950A2
公开(公告)日：2002-08-15
申请号：PCT/US2002/000247
申请日：2002-01-07
申请人： SBARRO INSTITUTE FOR CANCER RESEARCH AND MOLECULAR MEDICINE , CLAUDIO, Pier, Paolo
发明人： CLAUDIO, Pier, Paolo
IPC分类号： C12N
CPC分类号： G01N33/582 , C12N2799/022 , C12N2799/027 , C12Q1/70 , G01N33/57407 , C12Q2545/114
摘要： Titration is an important and critical step in dosing recombinant virus for gene therapy. A relatively fast, convenient and sensitive method that allows for precise quantification of recombinant retrovirus is presented. The method is based on PCR amplification of a foreign gene by the PRINS (primer in situ DNA synthesis) technique. The PRINS technique is based on the sequence-specific annealing of unlabeled oligonucleotide DNA in situ. This oligonucleotide operates as a primer for in situ chain elongation catalyzed by the Taq I polymerase. Using -labeled nucleotides as a substrate for chain elongation, the neo-synthetic DNA is labeled by an FITC-conjugated anti- antibody. To avoid the possibility of false positives, the puromycin resistance gene, which is associated with the transgene in the same viral vector and is not normally present in mammalian cells was amplified. The retroviral titer was evaluated by counting FITC-positive cells after PRINS labeling, while knowing the number of cells that were transduced with different amounts of viral supernatant. A comparable viral concentration of 1 X 10 7 infectious units/mL was found among the retroviruses.
摘要翻译：滴定是给予重组病毒进行基因治疗的重要和关键步骤。提出了一种相对快速，方便和灵敏的方法，可以对重组逆转录病毒进行精确定量。该方法基于PRINS（引物原位DNA合成）技术对外源基因的PCR扩增。 PRINS技术是基于原位未标记的寡核苷酸DNA的序列特异性退火。该寡核苷酸作为由Taq I聚合酶催化的原位链延长引物。使用标记的核苷酸作为链延长的底物，新合成的DNA用FITC缀合的抗体标记。为了避免假阳性的可能性，扩增了与同一病毒载体中的转基因相关并且通常不存在于哺乳动物细胞中的嘌呤霉素抗性基因。通过在PRINS标记后计数FITC阳性细胞来评估逆转录病毒滴度，同时知道用不同量的病毒上清转导的细胞数。在逆转录病毒中发现1×10 7感染单位/ mL的相当的病毒浓度。

2. 发明申请

WO2009117098A2 METHODS FOR STEM CELL PRODUCTION AND THERAPY 审中-公开
标题翻译：干细胞生产和治疗方法
公开(公告)号：WO2009117098A2
公开(公告)日：2009-09-24
申请号：PCT/US2009/001695
申请日：2009-03-18
申请人： MARSHALL UNIVERSITY RESEARCH CORPORATION , VALLURI, Jagan, V. , CLAUDIO, Pier, Paolo
发明人： VALLURI, Jagan, V. , CLAUDIO, Pier, Paolo
IPC分类号： C12N5/08
CPC分类号： C12N5/0695 , C12N2500/36 , C12N2525/00
摘要： The present invention relates to methods for rapidly expanding a stem cell population with or without culture supplements in simulated micro gravity conditions. The present invention relates to methods for rapidly increasing the life span of stem cell populations without culture supplements in simulated microgravity conditions. The present invention also relates to methods for increasing the sensitivity of cancer stem cells to chemo therapeutic agents by culturing the cancer stem cells under microgravity conditions and in the presence of omega-3 fatty acids. The methods of the present invention can also be used to proliferate cancer cells by culturing them in the presence of omega-3 fatty acids. The present invention also relates to methods for testing the sensitivity of cancer cells and cancer stem cells to chemotherapeutic agents by culturing the cancer cells and cancer stem cells under microgravity conditions. The methods of the present invention can also be used to produce tissue for use in transplantation by culturing stem cells or cancer stem cells under microgravity conditions. The methods of the present invention can also be used to produce cellular factors and growth factors by culturing stem cells or cancer stem cells under microgravity conditions. The methods of the present invention can also be used to produce cellular factors and growth factors to promote differentiation of cancer stem cells under microgravity conditions.
摘要翻译：本发明涉及用于在模拟微重力条件下快速扩张具有或不具有培养基的干细胞群体的方法。本发明涉及用于在模拟微重力条件下快速增加干细胞群体而没有培养补充剂的寿命的方法。本发明还涉及通过在微重力条件下和在ω-3脂肪酸存在下培养癌症干细胞来增加癌症干细胞对化疗治疗剂的敏感性的方法。本发明的方法也可用于通过在ω-3脂肪酸存在下培养它们来增殖癌细胞。本发明还涉及通过在微重力条件下培养癌细胞和癌干细胞来测试癌细胞和癌干细胞对化学治疗剂的敏感性的方法。本发明的方法还可用于通过在微重力条件下培养干细胞或癌干细胞来产生用于移植的组织。本发明的方法还可用于通过在微重力条件下培养干细胞或癌干细胞来产生细胞因子和生长因子。本发明的方法也可用于产生细胞因子和生长因子，以促进微重力条件下癌干细胞的分化。

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