Cell Culture

项目总结（见说明）：核心C 细胞培养将为本计划项目提供细胞培养支持。我们使用的检测方法可以检测出 造血祖细胞在不同的分化阶段，并可以区分正常和 白血病集落形成细胞这些分析非常适合解决本计划中出现的一个主要问题 项目，即衍生的细胞响应于各种抗白血病药物的调查， P01。这些试验将用于研究外源性细胞因子，包括粒细胞巨噬细胞的作用 集落刺激因子（GM-CSF）和c-Kit配体保护CML祖细胞免受伊马替尼的影响 (IM Gleevec）-使用对化疗敏感和耐药的患者的血液和骨髓细胞诱导细胞凋亡。 格列卫这些研究将与项目4和项目5合作进行。此外，我们将提供 支持项目5，研究脂质运载蛋白和反义24 p3和BCR对Ph+细胞系的影响， 正常和brc-abl+早期和成熟祖细胞。我们将与项目3合作，研究 破坏CML祖细胞的微环境保护将影响CML的敏感性 蛋白激酶抑制剂对集落形成细胞的影响，以及Bcl-2抑制剂是否会增强 蛋白激酶抑制剂。 因此，具体目标如下： 1)与Deininger博士（项目4）和Arlinghaus博士（项目5）合作，研究外源性刺激性细胞因子是否保护CML祖细胞免受IM和其他蛋白激酶抑制剂的凋亡作用 2)与Arlinghaus博士合作研究抗24 p3和抗NGAL抗体以及BCR（通过慢病毒载体感染系统转导）对CML细胞系和新鲜CML和正常骨髓细胞的影响（项目5）。 3)研究造血基质和CML克隆之间相互作用的中断如何影响正常和BCR-ABL+祖细胞对蛋白酪氨酸激酶抑制剂的敏感性。我们将与Andreeff博士（项目3）合作，使用造血基质培养物、长期培养起始细胞（LTCIC）试验和供体试验的组合，研究中断SDF-1和CXCR 4之间相互作用的药物的作用，以及酪氨酸激酶和Bcl-2抑制剂的组合。

PROJECT SUMMARY (See instructions): Core C The cell culture will provide cell culture support for this Program Project. The assays we use can detect hematopoietic progenitors at various stages of differentiation and can distinguish between normal and leukemic colony-forming cells. These assays are well suited to address a major issue arising in this Program Project, namely the derivation ofthe cells responding to various anti-leukemic agents investigated in this P01. These assays will be used to investigate the role of extrinsic cytokines including granulocytemacrophage colony stimulating factor (GM-CSF) and c-Kit ligand in protecting CML progenitors from Imatinib (IM; Gleevec)-induces apoptosis using blood and marrow cells of both patients sensitive and resistant to Gleevec. These studies will be conducted in collaboration with Projects 4 and 5. Also, we will provide support to Project 5 in studying the effects of lipocalin and antisense 24p3 and BCR on Ph+ cell lines and on normal and brc-abl+ eariy and mature progenitors. In collaboration with Project 3 we will study whether disruption ofthe microenvironment-dedicated protection of CML progenitors will affect the sensitivity of CML colony-forming cells to protein kinase inhibitors and whether Bcl-2 inhibitors will potentiate the effects of protein kinase inhibitors. Thus, the Specific Aims are the following: 1) To investigate whether extrinsic stimulatory cytokines protect CML progenitors from apoptotic effect of IM and other protein kinase inhibitors in collaboration with Drs. Deininger (Project 4) and Arlinghaus (Project 5) 2) To study the effects of anti-24p3 and anti-NGAL antibodies and BCR (transduced by the lentivirus vector infection system) on CML cell lines and fresh CML and normal marrow cells in collaboration with Dr. Arlinghaus (Project 5). 3) To investigate how interruption ofthe interaction between the hematopoietic stroma and the CML clone would affect the sensitivity of normal and BCR-ABL+ progenitors to protein tyrosine kinase inhibitors. We will study the effect of agents that interrupt the interaction between SDF-1 and CXCR4, and the combination of tyrosine kinases and Bcl-2 inhibitors using a combination of hematopoietic stroma cultures, the long-term culture initiating cell (LTCIC) assay and donogenic assays, in collaboration with Dr. Andreeff (Project 3).