HemaTox™ Megakaryocyte Kit

Serum-free medium and supplement for the measurement of drug toxicity on megakaryocytes
HemaTox™ Megakaryocyte Kit

Serum-free medium and supplement for the measurement of drug toxicity on megakaryocytes

1 Kit
Catalog # 09707
752 USD

Overview

Screen large numbers of novel drug compounds for their effect on the proliferation and megakaryocyte-specific differentiation of human hematopoietic stem and progenitor cells (HSPCs).

Compared to using the colony forming unit (CFU) assay, the liquid culture-based HemaTox™ assay is more flexible: you can add test compounds at various points throughout the culture and measure their effects on more primitive or more mature progenitor cells. The HemaTox™ Megakaryocyte Kit promotes the proliferation of human CD34+ HSPCs and their differentiation into CD41-expressing megakaryocytes.

Users can choose the readout method to quantify the response and estimate the IC50 and IC90 of each compound tested. Test up to 160 conditions—in triplicate—per kit in 5 x 96-well plates. This kit may be used on its own or in combination with HemaTox™ Erythroid Kit (Catalog #09701) or HemaTox™ Myeloid Kit (Catalog #09704)to assess lineage-specific drug toxicity in parallel.
Components
  • HemaTox™ Megakaryocyte Medium, 100 mL
  • HemaTox™ Megakaryocyte 100X Supplement, 1 mL
Subtype
Specialized Media
Cell Type
Hematopoietic Stem and Progenitor Cells, Megakaryocytes
Species
Human
Application
Cell Culture, Toxicity Assay
Brand
HemaTox
Area of Interest
Drug Discovery and Toxicity Testing, Stem Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
HemaTox™ Megakaryocyte Kit
Catalog #
09707
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
HemaTox™ Megakaryocyte Kit
Catalog #
09707
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
HemaTox™ Megakaryocyte Kit
Catalog #
09707
Lot #
All
Language
English

Educational Materials (4)

Brochure
Hematopoietic Stem and Progenitor Cells - Products for Your Research
Technical Bulletin
Media and Supplements for In Vitro Hematotoxicity Testing
Webinar
Anticipating Cytopenia: In Vitro Methods for Hematotoxicity Testing
55:33
Anticipating Cytopenia: In Vitro Methods for Hematotoxicity Testing
Scientific Poster
A Novel 96-well Plate Cell Culture Assay for Lineage-Specific Hematopoietic Cell Toxicity Screening

Data and Publications

Data

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.

Lineage-specific differences in hematotoxicity identified with HemaTox™ Erythroid, Myeloid and Megakaryocyte Kits

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).

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