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人和小鼠造血祖细胞的长期培养

MyeloCult™ 培养基专为造血祖细胞与贴壁基质细胞的长期共培养而开发。该培养基的重要应用在于通过 长期培养启动细胞(LTC-IC)检测 定量原始造血祖细胞。 在此检测中,将造血细胞培养在含有支持性人或小鼠基质细胞层的MyeloCult™培养基中,并通过后续使用 MethoCult™ 培养基进行的 集落形成单位(CFU)检测,评估其数周内生成髓系克隆形成祖细胞的能力。

为何使用 MyeloCult™ ?

  • 建立基质细胞层,实现"Dexter型"骨髓长期培养体系。1,2
  • 在MyeloCult™培养基中进行LTC-IC检测。3
  • 研究长期培养中骨髓造血作用的调节因子。3

MyeloCult™H5100

物种:

适用于以下体系的建立与维持:

  • 人造血细胞的髓系长期培养体系
  • 基质细胞饲养层(如M2-10B4小鼠骨髓成纤维细胞系)
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实验方案

图1. 有限稀释法长期培养启动细胞(LTC-IC)检测的流程



Procedure for Bulk Culture LTC-IC Assay

图2. 批量培养LTC-IC检测操作流程



Key Applications

Human Hematopoietic Cell Culture

Nemeth M et al. (2009) beta-Catenin expression in the bone marrow microenvironment is required for long-term maintenance of primitive hematopoietic cells. Stem Cells 27(5): 1109-1119
Tipping A et al. (2009) High GATA-2 expression inhibits human hematopoietic stem and progenitor cell function via effects on cell cycle. Blood
Peterson M et al. (2008) Inhibitory receptor signaling via tyrosine phosphorylation of the adaptor Crk. Immunity 29(4): 578-588
Carlo-Stella C et al. (2007) Placental growth factor-1 potentiates hematopoietic progenitor cell mobilization induced by granulocyte colony-stimulating factor in mice and nonhuman primates. Stem Cells 25(1): 252-261
Pastos K et al. (2006) Differential effects of recombinant thrombopoietin and bone marrow stromal-conditioned media on neonatal versus adult megakaryocytes. Blood 108(10): 3360-3362
Sloand E et al. (2006) Granulocyte colony-stimulating factor preferentially stimulates proliferation of monosomy 7 cells bearing the isoform IV receptor. Proc Natl Acad Sci 103(39): 14483-14488
Pirson L et al. (2006) Despite inhibition of hematopoietic progenitor cell growth in vitro, the tyrosine kinase inhibitor imatinib does not impair engraftment of human CD133+ cells into NOD/SCIDbeta2mNull mice. Stem Cells 24(7): 1814-1821
Takeda A et al. (2006) NUP98-HOXA9 induces long-term proliferation and blocks differentiation of primary human CD34+ hematopoietic cells. Cancer Res 66(13): 6628-6637
Giebel B et al. (2006) Primitive human hematopoietic cells give rise to differentially specified daughter cells upon their initial cell division. Blood 107(5): 2146-2152
Srour E et al. (2005) Modulation of in vitro proliferation kinetics and primitive hematopoietic potential of individual human CD34+CD38-/lo cells in G0. Blood 105(8): 3109-3116
Selleri C et al. (2005) Involvement of the urokinase-type plasminogen activator receptor in hematopoietic stem cell mobilization. Blood 105(5): 2198-2205
Giebel B et al. (2004) Segregation of lipid raft markers including CD133 in polarized human hematopoietic stem and progenitor cells. Blood 104(8): 2332-2338
Kogler G et al. (2004) A new human somatic stem cell from placental cord blood with intrinsic pluripotent differentiation potential. J of Exp Med 200(2): 123-135
Guan Y et al. (2003) Detection, isolation, and stimulation of quiescent primitive leukemic progenitor cells from patients with acute myeloid leukemia (AML). Blood 101(8): 3142-3149
Bartolovic K et al. (2004) Inhibitory effect of imatinib on normal progenitor cells in vitro. Blood 103(2): 523-529
Stier S et al. (2003) Ex vivo targeting of p21Cip1/Waf1 permits relative expansion of human hematopoietic stem cells. Blood 102(4): 1260-1266
Danet G et al. (2003) Expansion of human SCID-repopulating cells under hypoxic conditions. J of Clin Invest 112(1): 126-135
Dybedal I et al. (2003) Human reconstituting hematopoietic stem cells up-regulate Fas expression upon active cell cycling but remain resistant to Fas-induced suppression. Blood 102(1): 118-126
Punzel M et al. (2003) The symmetry of initial divisions of human hematopoietic progenitors is altered only by the cellular microenvironment. Exp Hematol 31(4): 339-347
Murdoch B et al. (2003) Wnt-5A augments repopulating capacity and primitive hematopoietic development of human blood stem cells in vivo. Proc Natl Acad Sci 100(6): 3422-3427
Kootstra N et al. (2003) Abrogation of postentry restriction of HIV-1-based lentiviral vector transduction in simian cells. Proc Natl Acad Sci U S A 100(3): 1298-1303
Bhatia M et al. (1998) A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat Med 4(9): 1038-1045
Dybedal I et al. (2003) Human reconstituting hematopoietic stem cells up-regulate Fas expression upon active cell cycling but remain resistant to Fas-induced suppression. Blood 102(1): 118-126

Mouse Hematopoietic Cell Culture

Zimmer S et al. (2011) Crebbp haploinsufficiency in mice alters the bone marrow microenvironment leading to loss of stem cells and excessive myelopoiesis. Blood
Lymperi S et al. (2010) Inhibition of osteoclast function reduces hematopoietic stem cell numbers in vivo. Blood 117(5): 1540-1549
Ohno Y et al. (2010) Hoxb4 transduction down-regulates Geminin protein, providing hematopoietic stem and progenitor cells with proliferation potential. Proc Natl Acad Sci
Ohmori T et al. (2010) Vinculin is indispensable for repopulation by hematopoietic stem cells, independent of integrin function.J Biol Chem 285(41): 31763-31773
Zhang Q et al. (2010) Fancd2-/- mice have hematopoietic defects that can be partially corrected by resveratrol. Blood
Sumitomo A et al. (2010) Transcriptional Mediator Subunit MED1/TRAP220 in Stromal Cells Is Involved in Hematopoietic Stem/Progenitor Cell Support through Osteopontin Expression. Mol Cell Biol
Pandit T et al. (2006) Sustained exposure to nicotine leads to extramedullary hematopoiesis in the spleen.Stem cells 24(11): 2373-2381
Lee Y et al. (2006) CCAAT/enhancer binding proteins alpha and epsilon cooperate with all-trans retinoic acid in therapy but differ in their antileukemic activities. Blood 108(7): 2416-2419
Carella C et al. (2006) The ETS factor TEL2 is a hematopoietic oncoprotein. Blood 107(3): 1124-1132
Carmen M et al. (2004) Interleukin-6 deficiency affects bone marrow stromal precursors, resulting in defective hematopoietic support.Blood 103(9): 3349-3354
Lacout C et al. (2003) A defect in hematopoietic stem cell migration explains the nonrandom X-chromosome inactivation in carriers of Wiskott-Aldrich syndrome. Blood 102(4): 1282-1289
Truong B et al. (2003) CCAAT/Enhancer binding proteins repress the leukemic phenotype of acute myeloid leukemia. Blood 101(3): 1141-1148
Jin H et al. (2002) Intracerebral transplantation of mesenchymal stem cells into acid sphingomyelinase-deficient mice delays the onset of neurological abnormalities and extends their life span. J of Clin Invest 109(9): 1183-1191
Samper E et al. (2002) Long-term repopulating ability of telomerase-deficient murine hematopoietic stem cells.Blood 99(8): 2767-2775
Iversen P et al. (2002) Decreased hematopoiesis in bone marrow of mice with congestive heart failure. Am J of Physiol, Regul, Integr Comp Physiol 282(1): R166-72
Jasinski M et al. (2001) GATA1-Cre mediates Piga gene inactivation in the erythroid/megakaryocytic lineage and leads to circulating red cells with a partial deficiency in glycosyl phosphatidylinositol-linked proteins (paroxysmal nocturnal hemoglobinuria type II cells). Blood 98(7): 2248-2255

参考文献

  1. Dexter T et al. (1977) Conditions controlling the proliferation of haematopoietic stem cells in vitro. J Cell Physiol 91(3): 335–44
  2. Gartner S et al. (1980) Long-term culture of human bone marrow cells. Proc Natl Acad Sci 77(8): 4756–9
  3. Sutherland H et al. (1990) Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci 87(9): 3584–8
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