The HALO®-96 SPC-QC Platform was specifically designed for stem and progenitor cell potency assays to accommodate different applications. CAMEO™-96 STD, the latest addition to these potency assays, uses the same 96-well plate configuration as the HALO®-96 SPC-QC assays and can be used for the same applications described below. Each of the configurations described are available in the HALO®-96 MeC, HALO®-96 SEC and CAMEO™-96 STD Platforms. There are 4 main assay kit configurations for each platform:
HALO®-96 SPC-QC and CAMEO™-96 STD Single Stem Cell Assay that detects the proliferation / growth potential of the mature multipotential CFC-GEMM population. This assay is comparable to the present traditional colony-forming assay procedure performed in stem cell processing laboratories. It is ideal for small to very large sample numbers. A total of 16 samples, each using 6 x 100µl replicates, can be assayed on a single plate, but do not have to be assayed all at the same time. This kit is available in 1, 2, 3 and 4 plate sizes.
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Download the HALO® SPC-QC Single Stem Cell Protocol
HALO®-96 SPC-QC and CAMEO™-96 STD Duel Stem Cell Assay. This kits allows the proliferation / growth potential of 2 types of stem cells, the primitive High Proliferative Potential Stem and Progenitor Cell (HPP-SP) and the mature multipotential CFC-GEMM. The HPP-SP cell population is positioned approximately at the point where lymphopoiesis and hematopoiesis diverge into 2 systems. The HPP-SP therefore detects primitive stem cells with high proliferative capacity. Their presence also implies the presence of more primitive stem cell populations such as the Long-Term Culture - Initiating Cell (LTC-IC). By assaying for both the HPP-SP and CFC-GEMM populations, it might be possible to implicate the presence of both short- and long-term engrafting and repopulating cells. A total of 8 samples using 6 x 100µl replicates , can be assayed on a single plate. This kit is also available in 1, 2, 3 and 4 plate sizes.
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Download the HALO® SPC-QC Duel Stem Cell protocol
HALO®-96 SPC-QC and CAMEO™-96 STD 4-Population Assay. When a CFC-GEMM traditional colony-forming assay is performed, the combination of growth factors allows the stimulation of the multipotential stem cell. However, the growth factors and cytokines also stimulate BFU-E, GM-CFC and Mk-CFC. However, it is not possible to distinguish whether the presence of these lineage-specific populations were derived from the stem cell population or whether they were present in the processed stem cell product. The HALO® SPC-QC 4-Population assay detects, CFC-GEMM, BFU-E, GM-CFC and Mk-CFC as separate, independent cell populations. As a result, the presence and proliferative potential of each cell population can be ascertained individually, but simultaneously in side-by-side cultures. The advantage of this is that it might provide information on short-term engraftment and repopulation potential of the sample. This kit can be used to measure 4 samples, each 6 x 100µl replicates. These kits are also available in 4 plates sizes.
HALO®-96 SPC-QC and CAMEO™-96 STD 7-Population Assay. The 7-population system is a combination of the HALO® Duel Stem Cell and the HALO® 4-Population Kit. However, also included are 2 lymphopoietic cell populations and a background control. Thus, the 7-Population Assay includes detection and measurement of HPP-SP, CFC-GEMM, BFU-E, GM-CFC, Mk-CFC, T-CFC and B-CFC. These populations, together with the background control (no growth factors) are assayed side-by-side as 2 samples on a single 96-well plate using 6 x 100µl replicate cultures. The graphs below show how the HALO® SPC-QC 7-Population Platform can be used for both human bone marrow and human umbilical cord blood.
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Download the HALO® SPC-QC 7-Population Protocol
This assay system could provide the most information that might predict both long- and short-term engraftment and reconstitution of the lympho-hematopoietic system. The HALO® SPC-QC 7-Population Assay has been developed into the HALO® 7-Population Predictive Hemotoxicity Platform, which has been validated against the Registry of Cytotoxicity Prediction Model. This proprietary toxicity testing platform has been shown not only to predict the response of the lympho-hematopoietic system to numerous agents, but also clinically-relevant dosing of anti-cancer drugs to human patients.
HALO®-96 SPC-QC Expansion Potential Kit
The HALO® SPC-QC Expansion Potential Kit was specifically designed to determine the presence of quiescent primitive high proliferative potential stem and progenitor cells (HPP-SP) and their potential for expansion into multiple lympho-hematopoietic lineages. The HPP-SP stem cell population tested in the Dual and 7-Population SPC-QC assays are “fully stimulated” with a combination of growth factors that allows the cells to enter multiple lineages of the lympho-hematopoietic system.
The HALO® SPC-QC Expansion Potential protocol takes this one step further by culturing the HPP-SP in a 2-stage culture system. The majority of the HPP-SP are in a quiescent state, that is, the cells are out of cell cycle. In the first stage of culture, the cells are “primed” out of quiescence and into cell cycle over a 5-7 day period. A portion of the replicate cultures are used to determine the proliferative potential of the cells by bioluminescence. Extra replicate cultures can be prepared so that after this primary culture, cells can be removed for phenotypic analysis or other assays. The cells from the remaining replicate cultures are removed from the wells and re-plated into a secondary culture containing a combination of growth factors and cytokines that allows full stimulation into multiple lympho-hematopoietic lineages. The secondary culture is incubated for a further 7 days and a portion of the replicate wells used to determine the proliferative potential by luminescence. Cells in remaining cultures can be removed and used for phenotypic analysis, gene expression profiling, etc.
The HALO® SPC-QC Expansion Potential protocol can be used for mononuclear cells or purified cells from bone marrow, peripheral blood (normal and mobilized) and umbilical cord blood. Since the protocol requires a secondary re-plate of the “primed” HPP-SP cells, it provides the substantiation that these primitive stem cells were present in the original sample. The ability of these cells to be induced into cell cycle and exhibit expansion might be important in predicting the role these cells might play during transplantation.
The HALO® SPC-QC Expansion Potential Kit is available as a 2 or 4 plate kit.
LINK
Download the HALO® SPC-QC Expansion Potential Protocol
