HemaTox™ Megakaryocyte Kit for In Vitro Toxicity Assays

总览

该产品为筛选大量新药化合物对人造血干祖细胞（HSPC）增殖和往巨核细胞定向分化的影响而设计。

与使用集落形成单位（CFU）实验相比，基于液体培养的HemaTox™实验更加灵活：您可以在整个培养过程的不同时间点添加测试化合物，测定其对更原始或更成熟的祖细胞的影响。HemaTox™巨核细胞试剂盒促进CD34+ HSPC增殖以及分化为表达CD41的巨核细胞。

用户可以自选读出方法来量化细胞对每个待测化合物的反应以及估计化合物的IC50和IC90。每套试剂盒可在5块96孔板中测试多达160个条件（三复孔）。该试剂盒可单独使用或与HemaTox™红系试剂盒（产品号 #09701）或HemaTox™髓系试剂盒（产品号 #09704）联合使用，以平行评估谱系特异性药物造血毒性。

亚型
专用培养基

细胞类型
造血干/祖细胞，巨核细胞

种属
人

应用
细胞培养，细胞毒性检测

品牌
HemaTox

研究领域
药物发现和毒理检测，干细胞生物学

制剂类别
无血清

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Data Figures

General HemaTox™ Erythroid Kit Procedure

Figure 1. General HemaTox™ Megakaryocyte Kit Procedure

*The cell isolation step may be omitted if pre-enriched CD34+ cells are used.

Flow cytometry plots showing erythroid cells produced after culture of CD34+ HSPCs with the HemaTox™ Erythroid Kit

Figure 2. Flow Cytometry Plot Showing Cells Produced After Culture of CD34+ HSPCs with the HemaTox™ Megakaryocyte Kit

(A) Human CB CD34+ cells were cultured with the HemaTox™ Megakaryocyte Kit using the protocol as written in the Product Information Sheet (PIS). (B) After the appropriate culture period, cells were harvested and stained for cell surface proteins expressed on megakaryocyte (CD41 and CD45) cells.

Reproducibility of HemaTox™ Erythroid Kit results between experiments and using different CD34+ cell preparations

Figure 3. Reproducibility of HemaTox™ Megakaryocyte Kit Results Between Experiments and Using Different CD34+ Cell Preparations

(A) Dose-response curves were generated from titrations of 5-FU added to human CB CD34+ cells isolated from 3 donors and cultured with the HemaTox™ Megakaryocyte Kit. Three to five separate experiments were performed with cells from each donor. In each assay similar IC50 values were obtained with cells from different donors and in different experiments with cells from the same donor. Shown are values normalized to the percentages (%) of maximum cell growth without drug. Despite differences in absolute cell counts, curves are reproducible when normalized within each experiment. (B) Table showing IC50 values generated for 5-FU in culture with the megakaryocyte kit including standard deviation (SD) and the coefficient of variation (% CV) across experiments.

Figure 4. Lineage-Specific Differences in Hematotoxicity Identified with HemaTox™ Erythroid, Myeloid and Megakaryocyte Kits

Results show average IC50 values for each drug tested on human BM CD34+ cells using the HemaTox™ Erythroid (grey), Myeloid (gold) and Megakaryocyte (orange) Kits. Most drugs show similar toxicity for each lineage but some, such as Sunitinib, are ~100-fold more toxic for erythroid than for megakaryocyte progenitor differentiation with intermediate toxicity for myeloid progenitor differentiation. Vertical lines indicate standard error of the mean (SEM) (n = 4 - 8).