EasySep™小鼠TIL（CD45）正选试剂盒
MegaCult™ 培养基用于 CFU-Mk 检测
巨核细胞祖细胞集落测定的胶原基培养基
megadult™-C是一种基于胶原蛋白的培养系统，用于人源或鼠源造血组织巨核细胞祖细胞（CFU- mk）的集落形成单位（CFU）检测。1仅通过形态学标准难以检测巨核细胞集落，因为集落可能仅包含三个细胞，且巨核细胞在形态上不易与其他细胞谱系（特别是巨噬细胞）区分。与其他谱系CFU检测中使用的甲基纤维素培养基不同，MegaCult™中的胶原凝胶可轻松脱水，便于后续进行细胞化学或免疫细胞化学染色。在MegaCult™中，人源CFU-Mk集落的检测基于对巨核细胞标记物CD41的免疫细胞化学染色，而鼠源集落检测则基于乙酰胆碱酯酶活性。
MegaCult™培养基提供含细胞因子/脂质与不含细胞因子/脂质两种规格。含细胞因子的MegaCult™培养基包含重组人(rh)IL-3、IL-6和TPO，可优化人源CFU-Mk的生长。当需要控制细胞因子类型和用量时（例如研究影响巨核祖细胞生长和成熟的因素），推荐使用不含细胞因子的MegaCult™培养基。不含细胞因子的MegaCult™-C培养基也用于小鼠CFU-Mk的培养和计数，添加适当的细胞因子，如重组细胞 鼠IL-3, rhIL-6, rhIL-11, rhTPO 和/或用户选择的其他因子。
MegaCult™-C
完整试剂盒
规格(培养基):
- 24 × 2ml 管（含细胞因子）
- 24 支1.7 mL 试管（仅限培养基）
物种:
人
细胞因子:
有或没有重组细胞因子
包含:
- 胶原蛋白溶液
- 培养基
- CFU-Mk染色试剂盒
- 双腔细胞培养玻片
应用:
脐带血、骨髓和外周血巨核细胞祖细胞的培养和计数。
MegaCult™-C
胶原蛋白和培养基
规格(媒体):
- 24 × 2ml 管（含细胞因子）
- 24 支1.7 mL 试管（仅限培养基）
物种:
- 人
- 小鼠
细胞因子:
可选已添加重组细胞因子（仅限人）或未添加重组细胞因子（人或小鼠）
包含:
- 胶原蛋白溶液
- 培养基
应用:
- 人脐带血、骨髓和外周血的 CFU-Mk 测定。
- 小鼠骨髓的CFU-Mk测定。需要添加细胞因子。
MegaCult™-C
胶原蛋白和含脂培养基
规格(培养基):
50 mL 瓶
物种:
- 人
- 小鼠
细胞因子:
没有添加细胞因子
包含:
- 胶原蛋白溶液
- 含脂培养基
应用:
- 人脐带血、骨髓和外周血富集祖细胞制剂的CFU-Mk测定。需要添加细胞因子。
- 小鼠骨髓的CFU-Mk测定。需要添加细胞因子。
- 在添加用户选择的适当细胞因子后，CFU检测其他祖细胞。
收起详情
数据
图1. 源自人巨核细胞祖细胞的中小型CFU-Mk菌落的例子
操作步骤
图2. 人巨核细胞祖细胞测定方法
图3. 小鼠巨核细胞祖细胞测定方法
Key Applications
Drug Toxicity Testing
Pessina A et al. (2009) Application of human CFU-Mk assay to predict potential thrombocytotoxicity of drugs. Toxicol In Vitro 23(1): 194-200
Transplantation and Engraftment Kinetics of Human HSPCs
Yasui K et al. (2003) Differences between peripheral blood and cord blood in the kinetics of lineage-restricted hematopoietic cells: implications for delayed platelet recovery following cord blood transplantation. Stem Cells 21(2): 143-151
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
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
Megakaryocyte Biology
Matsumura-Takeda K et al. (2007) CD41+/CD45+ cells without acetylcholinesterase activity are immature and a major megakaryocytic population in murine bone marrow. Stem Cells 25(4): 862-870
Boyer L et al. (2008) Increased production of megakaryocytes near purity from cord blood CD34+ cells using a short two-phase culture system. J Immunol Methods 332(1-2): 82-91
Vodyanik M et al. (2006) Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures. Blood 108(6): 2095-2105821
Boyer L et al. (2008) Increased production of megakaryocytes near purity from cord blood CD34+ cells using a short two-phase culture system. J Immunol Methods 332(1-2): 82-91
Vodyanik M et al. (2006) Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures. Blood 108(6): 2095-2105821
HSPC Biology
Goossens S et al. (2011) The EMT regulator Zeb2/Sip1 is essential for murine embryonic hematopoietic stem/progenitor cell differentiation and mobilization. Blood 117(21): 5620-5630
Migliaccio A et al. (2003) GATA-1 as a regulator of mast cell differentiation revealed by the phenotype of the GATA-1low mouse mutant. J Exp Med 197(3): 281-296
Ito C et al. (2003) Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice.
Blood 101(2): 517-523
Wang Y et al. (2011) Fine-tuning p53 activity through C-terminal modification significantly contributes to HSC homeostasis and mouse radiosensitivity. Genes Dev 25(13): 1426-1438
Lannutti B et al. (2008) Incomplete restoration of Mpl expression in the mpl-/- mouse produces partial correction of the stem cell repopulating defect and paradoxical thrombocytosis. Blood 113(8): 1778-1785
Hisa T et al. (2004) Hematopoietic, angiogenic and eye defects in Meis1 mutant animals. EMBO J 23(2): 450-459
Xing S et al. (2008) Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice. Blood 111(10): 5109-5117
Wernig G et al. (2006) Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model. Blood 107(11): 4274-4281
Moody J et al. (2004) Anemia, thrombocytopenia, leukocytosis, extramedullary hematopoiesis, and impaired progenitor function in Pten+/-SHIP-/- mice: a novel model of myelodysplasia. Blood 103(12): 4503-4510
Migliaccio A et al. (2003) GATA-1 as a regulator of mast cell differentiation revealed by the phenotype of the GATA-1low mouse mutant. J Exp Med 197(3): 281-296
Ito C et al. (2003) Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice.
Blood 101(2): 517-523
Wang Y et al. (2011) Fine-tuning p53 activity through C-terminal modification significantly contributes to HSC homeostasis and mouse radiosensitivity. Genes Dev 25(13): 1426-1438
Lannutti B et al. (2008) Incomplete restoration of Mpl expression in the mpl-/- mouse produces partial correction of the stem cell repopulating defect and paradoxical thrombocytosis. Blood 113(8): 1778-1785
Hisa T et al. (2004) Hematopoietic, angiogenic and eye defects in Meis1 mutant animals. EMBO J 23(2): 450-459
Xing S et al. (2008) Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice. Blood 111(10): 5109-5117
Wernig G et al. (2006) Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model. Blood 107(11): 4274-4281
Moody J et al. (2004) Anemia, thrombocytopenia, leukocytosis, extramedullary hematopoiesis, and impaired progenitor function in Pten+/-SHIP-/- mice: a novel model of myelodysplasia. Blood 103(12): 4503-4510
参考文献
- Hogge D et al. (1997) Quantitation and characterization of human megakaryocyte colony-forming cells using a standardized serum-free agarose assay. Br J Hematol. 96(4): 790-800.
