DEVELOPMENT OF HEMATOPOIETIC STEM CELLS
造血干细胞的发育
基本信息
- 批准号:6383644
- 负责人:
- 金额:$ 25.23万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2001
- 资助国家:美国
- 起止时间:2001-07-01 至 2005-06-30
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Despite recent advances in understanding of mammalian embryonic hematopoiesis, site of origin and developmental potential of embryonic hematopoietic stem cells (HSC) remain unclear. Furthermore, recent observation that neuronal stem cells could have a much broader developmental capacity than anticipated, calls for reassessment of the full developmental potential of other tissue-specific stem cell types (e.g. HSC, mesenchymal, hepatic, epidermal stem cells etc.). New in utero transplantation assay termed blastocyst engraftment assay (BEA) will be used to further study embryonic hematopoiesis. BEA is based on microinjection of purified HSC into mouse preimplantation embryos (blastocysts), similar to ES cell technology. This new assay does not interfere with embryo development and results in engraftment of donor HSC in embryonic and adult hematopoietic tissues of recipients. The advantage of BEA is that the trafficking of donor HSC takes place during normal embryonic development, and secondly, the engraftment of donor HSC occurs through competitive repopulation of newly forming cell niches. The overall goal of the proposed project is to (a) study homing and engraftment of microinjected transgenic fetal and adult HSC during mouse embryo development in vivo and in vitro, and (b) comparatively analyze the potential of mouse fetal and adult HSC for development into non-hematopoietic cells/tissues. Towards this goal, the first part of the proposal focuses on the extensive in vivo analysis of the fate of transgenic fetal and adult HSC (isolated from ROSAbeta-geo26 and GFP transgenic mice) after microinjection into embryos of wild type and GATA-2 and SCL knockout mice. Engraftment of donor HSC in embryonic hematopoietic tissues will be assessed by analysis of embryos, tissues and cells using histochemical X-gal staining, fluorescence microscopy, histological and molecular techniques and in vitro hematopoietic colony forming assays (CFU-C). To complement in vivo studies, the second part deals with in vitro analysis of the fate of microinjected transgenic HSC. Culture of whole mouse embryos (reaching developmental stage of day 10 embryos in vivo) will be used as an experimental system for real time tracking of GFP-expressing transgenic HSC within the same embryos over the course of culture. The third part encompasses in vivo analysis of the potential of fetal and adult mouse HSC to differentiate into non-hematopoietic cells during mouse embryo development. The outcome of proposed studies could have important impact on the fields of embryonic hematopoiesis and clinical in utero and adult transplantation of HSC, .and perhaps on the development of new therapeutic approaches using autologous HSC to treat disorders or injuries affecting non-hematopoietic, tissues where the specific stem cells are not readily available for isolation.
尽管最近对哺乳动物胚胎造血的了解取得了进展,但胚胎造血干细胞(HSC)的起源和发育潜力仍不清楚。此外,最近的观察表明,神经干细胞的发育能力可能比预期的要广泛得多,这要求重新评估其他组织特异性干细胞类型(如HSC、间充质干细胞、肝脏干细胞、表皮干细胞等)的全部发育潜力。一种名为胚泡植入试验(BEA)的新的宫内移植试验将被用于进一步研究胚胎造血。BEA是基于将纯化的HSC显微注射到小鼠植入前胚胎(囊胚)中,类似于ES细胞技术。这种新的方法不会干扰胚胎发育,并导致供者的HSC植入受体的胚胎和成人造血组织中。BEA的优点是供体HSC的转运发生在正常胚胎发育期间,其次,供体HSC的植入是通过新形成的细胞壁龛的竞争性再繁殖来进行的。该项目的总体目标是(A)研究显微注射转基因胎儿和成人HSC在体内和体外胚胎发育过程中的归巢和植入,以及(B)比较分析小鼠胎儿和成人HSC向非造血细胞/组织发育的潜力。为了实现这一目标,该提案的第一部分侧重于对转基因胎儿和成年HSC(从ROSAbeta-ge26和GFP转基因小鼠中分离出来)在显微注射到野生型和GATA-2和SCL基因敲除小鼠的胚胎后的命运进行广泛的体内分析。供体HSC在胚胎造血组织中的植入将通过组织化学X-Gal染色、荧光显微镜、组织学和分子技术以及体外造血细胞集落形成试验(CFU-C)对胚胎、组织和细胞进行分析。为了补充体内研究,第二部分对显微注射转基因HSC的体外命运进行了分析。小鼠胚胎培养(体内发育到第10天胚胎)将作为一个实验系统,在培养过程中实时跟踪同一胚胎中表达GFP的转基因HSC。第三部分包括体内分析胎儿和成年小鼠的HSC在小鼠胚胎发育过程中分化为非造血细胞的潜力。建议的研究结果可能会对胚胎造血和临床子宫内和成人造血干细胞移植领域产生重要影响,也可能对开发新的治疗方法,使用自体骨髓干细胞来治疗影响非造血组织的疾病或损伤,这些组织中的特定干细胞不容易分离。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Roland Jurecic其他文献
Roland Jurecic的其他文献
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{{ truncateString('Roland Jurecic', 18)}}的其他基金
(PQ9) Characterization and prophylactic treatment of chemotherapy-induced long-term adverse sequelae
(PQ9)化疗引起的长期不良后遗症的特征和预防性治疗
- 批准号:
10000855 - 财政年份:2016
- 资助金额:
$ 25.23万 - 项目类别:
(PQ9) Characterization and prophylactic treatment of chemotherapy-induced long-term adverse sequelae
(PQ9)化疗引起的长期不良后遗症的特征和预防性治疗
- 批准号:
9752495 - 财政年份:2016
- 资助金额:
$ 25.23万 - 项目类别:
(PQ9) Characterization and prophylactic treatment of chemotherapy-induced long-term adverse sequelae
(PQ9)化疗引起的长期不良后遗症的特征和预防性治疗
- 批准号:
9547339 - 财政年份:2016
- 资助金额:
$ 25.23万 - 项目类别:
THE ROLE OF TR-KIT RECEPTOR IN STEM CELL MAINTENANCE AND DIFFERENTIATION
TR-Kit 受体在干细胞维持和分化中的作用
- 批准号:
7149857 - 财政年份:2006
- 资助金额:
$ 25.23万 - 项目类别:
THE ROLE OF TR-KIT RECEPTOR IN STEM CELL MAINTENANCE AND DIFFERENTIATION
TR-Kit 受体在干细胞维持和分化中的作用
- 批准号:
7268018 - 财政年份:2006
- 资助金额:
$ 25.23万 - 项目类别: