Introducing
ProliferStem™
A simple cellular functionality and viability assay performed in 30 minutes to predict proliferation potential of virtually any cell type.
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ProliferStem™ measures cellular functionality by measuring cellular and mitochondrial integrity.
All cells produce ATP. Intracellular ATP (iATP) is an excellent biochemical marker for several cell functions. It can be used to measure proliferation or cytotoxicity (as in the case of HALO®, LUMENESC™ and LumiSTEM™) and cell number. Intracellular ATP also detects cellular and mitochondrial integrity (CMI) and therefore cell viability. If cellular and mitochondrial integrity are intact, then cell functionality will also be intact. ProliferStem™ detects cell functionality.
The iATP concentration directly correlates with cell functionality and viability. Other viability markers that use dye exclusions, e.g. trypan blue, 7-aminoactinomycin D (7-AAD), acridine orange, only detect membrane integrity. They provide no indication regarding the integrity of the cellular and mitochondrial functions that are a prerequisite for healthy cell function. ProliferStem™ measures cell functionality by measuring the iATP concentration using one of the most sensitive signal detection readouts available, luciferin/luciferase-produced bioluminescence. If little or no ATP is produced, the cells will not proliferate or differentiate. Only when the cell can produce sufficient iATP will it have the capability to enter the proliferation process.
A Special Problem is found in stem cell transplantation for hematologic diseases. In this case, bone marrow and more recently, mobilized peripheral blood and umbilical cord blood are used as stem cell transplantation products. Umbilical cord blood, in particular, is cryopreserved for months or years prior to its use as a transplant product. ProliferStem™ can provide a rapid and easy assessment of whether a cord blood sample/unit will exhibit the adequate cell functionality to proliferate, a prerequisite for engraftment.
When to Use ProliferStem™
ProliferStem is used in place of, or in addition to, any dye exclusion viability assay to determine CMI and the ability to proliferate. ProliferStem should be used at the same time as total or mononuclear cell counts are determined and together with membrane or other intracellular expression markers.
ProliferStem™ Can Be Used With Virtually Any Cell Type.
ProliferStem™ can be used for any cell types for which cell functionality and proliferative potential is required prior to in vitro culture or other treatments.
A. Hematopoietic Cells: The diagram shows that ProliferStem can predict human bone marrow stem and progenitor cell proliferation when combined with HALO®-96 SEC. ProliferStem can be used in a similar manner with mobilized peripheral blood and umbilical cord blood. Also use ProliferStem™ for all animal hematopoietic cells (bone marrow, spleen, fetal liver, yolk sac) from different species.
B. Mesenchymal Stem Cells (MSC): Use ProliferStem™ prior to culturing MSC and measure their proliferation using LUMENESC™.
C. All Other Cell Types: ProliferStem™ can be used to determine the cellular functionality of ES cells, cell lines, ex vivo tissues of different species and more. Use ProliferStem™ prior to LumiSTEM™.
HOW ProliferStem™ Works
ProliferStem™ is a proprietary assay developed by HemoGenix®®. ProliferStem™ is based on the cell’s ability to produce intracellular ATP (iATP). If iATP is produced, the cell exhibits cellular and mitochondrial functionality. NO CELL CULTURE IS NECESSARY. Just dispense the cells concentrations into the 96-well plate provides and add 100µl of the ATP Monitoring Reagent. Wait 10 minutes. Intracellular ATP is released from cells and prodices a "glow" bioluminescence in the form of light in the presence of luciferin/luciferase. The amount of light (photons) is measured in a plate luminometer. The assay is calibrated and standardized against an external ATP standard. Healthy cells produce iATP concentrations of >0.05µM at 5,000 cells/well.
The assay takes only 30 minutes.
A Complete Family of ATP Bioluminescence Proliferation Assays.
ProliferStem™ belongs to a family of ATP bioluminescence assays developed by HemoGenix®. HALO® was the first platform to be developed and is based on and equivalent to the colony-forming cell (CFC) assay for lympho-hematopoietic cells. The LUMENESC™ Platform, specifically designed to detect Mesenchymal Stem Cells (MSC) was the second ATP-based proliferation assay developed. Finally, LumiSTEM™ can be used to detect and measure proliferation in virtually any other cell type, including ES cells cell lines and numerous expanted cells from tissues.