Stem cell hemotoxicity testing using the HALO® platform

by Alexey Bersenev on July 14, 2010 · 0 comments

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HemoGenix®, a Colorado based biotechnology firm specializes in stem cell hemotoxicity testing and screening and produces and manufactures assay kits for basic research, stem cell transplantation and cord blood banks and drug development and environmental testing.

HemoGenix® is the leader in in vitro, non-invasive, cell-based predictive and validated hemotoxicity testing using the HALO® platform. HALO® is based on the following principle: When lympho-hematopoietic cells proliferate or are inhibited from proliferating, the intracellular ATP (iATP) concentration changes proportionately. After incubation, the iATP is released from the cells by lysis. The iATP acts as a limiting substrate for a luciferin / luciferase reaction to produce bioluminescence in the form of light. The amount of light is a direct indication of the proliferation status of the cells.

The bioluminescence produced is detected in a plate luminometer. The luminometer readout is in non-standardized Relative Luminescence Units (RLU). To convert RLU into a standardized ATP unit, a standard ATP dose response is performed prior to measuring the samples. The software used to control the plate luminometer can usually be programmed to automatically convert RLU values into calibrated ATP (µM) values.

By calibrating your results to the external ATP standard provided with all luminescence kits, you can compare results directly from one experiment to another, regardless of the time span between experiments. Furthermore, you can also compare results between laboratories, thereby providing information to validate the assay for specific applications.

Essentially, the HALO® platform is a replacement for the traditional colony-forming cell (CFC) assay. Instead of manually counting differentiated clusters or colonies, the non-subjective and fully calibrated HALO® platform measures proliferation. Since proliferation and not differentiation, the status of lympho-hematopoietic cell populations can be measured within 4-7 days. HALO® is therefore more rapid and easier to use than the CFC assay. In addition, the actual time to process and read a full 96-or 384-well plate kit is only about 30 minutes or less.

HALO® can be used to detect and measure the proliferation of at least 12 different lympho-hematopoietic stem, progenitor and precursor cell populations from human, non-human primate, horse, sheep, dog, rat and mouse. The HALO® kits come complete with everything required to perform the assay except for the cells. A plate luminometer is also required.

Besides its easy of use, other major benefits of using HALO® is that manual colony counting is not necessary and the assay requires very little training and, unlike the CFC assay, HALO® can be validated to comply with regulatory requirements.

HemoGenix® also offers the LUMENESC assay specific for Mesenchymal Stem Cells and LumiSTEM, an assay available for ES, iPS, Primary Cells and Cell Lines. To learn more about these assays and the HemoGenix® patented technology, please visit www.hemogenix.com.

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