7.造血干细胞表型及分离纯化方法课件.ppt

抗Sca-1抗体经常和一些细胞表面标志分子联用一起用来鉴定和分离小鼠HSCs。Sca-1+ HSCs可在成年骨髓、胎儿肝脏、成年动物外周血和脾脏中被找到。Sca-1在几种非造血组织中也被发现。43 Sca-1可用来富集HSCs之外的祖细胞。Sca-1 可能参与调节B细胞和T细胞活化。 根据Sca-1是否阴性可将Thy-1lowLin-细胞进一步分为：Thy-1lowLin- Sca-1+ 和Thy-1lowLin- Sca-1-两群，前者含干细胞和CFU-T，后者不含 CFU-T，两群细胞都含CFU-s。 mouse 和 rat 骨髓造血干细胞中与淋巴系组细胞定向、分化有关的抗原。 2. Thy-1(CD90) 一种原癌基因。最近证明造血干细胞与c-kit基因密切相关。c-kit可编码一种穿膜酪氨酸激酶受体分子。应用单克隆抗体证明此分子可存在于造血干细胞膜上，其后证明它的配体分子是造血干细胞因子（stem cell factor, SCF），一种信号传导分子，对造血干细胞的分化具有重要作用。 3. c-kit (CD117) 　 c-kit分子可高频率表达于多能干细胞表面,但骨髓中c-kit+细胞可分化为各种血细胞，而胸腺中c-kit细胞可分化为淋巴细胞，不能分化为髓系细胞，所以胸腺内 c-kit+细胞，可能是淋巴样干细胞。 Lineage negative,指 T-、B-、M-、G- 细胞。 4. Lin- 97kDa的糖蛋白，包括5个跨膜结构域， 糖基化后在蛋白胶上显示120 kDa的蛋白带。最初是通过AC133单抗鉴定的，它能识别人HSCs的CD34+亚类29,30。一种CD133异构体AC133-2, 最近已经被克隆并鉴定为可被AC133抗体识别的原始表面抗原。 5. CD133(AC133) Chromosomal Location 4p16.2-p12 [human] The AC133 antigen is likely to contain 5-TM domains in addition to a signal peptide. The 5-TM structure includes an extracellular N-terminus, two short intracellular loops, two large extracellular loops and an intracellular C-terminus. Protein Sequence hematopoietic stem and progenitor cells. neural and embryonic stem cells. some leukemias and brain tumours that may be derived from stem cells. Low level expression could also be detected in the kidney, pancreas, placenta, and fetal liver. Expression MACS-isolated CD133+ cells became adherent and CD133- during cultivation, but gave rise to non-adherent CD133+ cells budding from the adherent cell surface by a symmetric cell division. Cells were stained with CD133/1 (AC133)-PE. CD133-expressing cells in different hematopoietic tissues 75.3 ± 0.8 1.75 ± 0.31 1.37 ± 0.27 Apheresis (采血) 51.0 ± 1.6 0.37 ± 0.06 0.16 ± 0.03 Cord blood 36.3 ± 2.2 1.47 ± 0.23 0.52 ± 0.11 Normal bone marrow CD133+ among CD34+ cells % CD34+ % CD133+ % ? Tree of differentiation 6. ABCG2? (ATP-binding cassette, sub-family G, member 2)?? Synonyms: ABC15, ABCP, BCRP, BCRP1