HALO®-96 SPC-QC

Stem and Progenitor Cell Quality Control Assays for the Stem Cell Processing Laboratory

Buy HALO®-96 SPC-QC assay kits

Available as assay kits and
contract research services.

Stem Cell "Quality" and Stem Cell Potency

Stem cell "quality" is the ability of stem cells to proliferate.
Stem cell "quality" cannot be measured using a CFU assay, which detects the ability of stem cells to differentiate and mature (see HALO® and CFC Assays).
Stem cell "quality" does not measure stem cell potency. However, a stem cell potency assay can measure stem cell "quality" (see HALO®-96 PQR, Potency, Quality, Release).

HALO®-96 SPC-QC Applications

For quantifying the "quality" of stem cells in a stem cell product prior to use.
To determine whether a processing procedure affects stem cell "quality".
To determine stem cell "quality" pre- and post cryopreservation.
To determine stem cell processing consistency and stability.

NOTE: Although the community norm is to use the colony-forming unit (CFU) assay, such as CAMEO™-4, an equivalent and alternative assay, such as HALO®-96 SPC-QC is acceptable by the FDA, since it satisfies "the requirements of the applicable statutes and regulations". In addition, AABB Standards for Cellular Therapy Services also allows "equivalent functional assays for any product", and the National Marrow Donor Program (NMDP) allows testing "consistent with AABB Standards".

Advantages to using HALO®-96 SPC-QC in the Stem Cell Processing Laboratory

HALO®-96 SPC-QC quantitatively measures proliferation ability and cellular functionality that cannot be enumerated using total nucleated counts (TNC), viability or flow cytometric detection of membrane expression markers.
HALO®-96 SPC-QC determines the "quality" of mature, multipotential hematopoietic stem cells (CFC-GEMM) and more primitive lympho-hematopoietic stem cells (HPP-SP).
Instrument-based, incorporating an external standard and controls for assay standardization and validation according to FDA regulations.
Compare results over time for intra- and inter-laboratory comparison studies.
Assay can be completed on day 5, 6 or 7 of culture (see below).
An FDA and AABB alternative "quality" assay.
Faster, easier to use and more accurate than any traditional CFU assay, even using automated colony counting.
HALO®-96 SPC-QC utilizes Suspension Expansion Culture (SEC) and bioluminomics™ Technology, the most advanced and cost-effective lympho-hematopoietic in vitro assay system available for the stem cell processing laboratory.

Tissues

Mobilized peripheral blood
Umbilical cord blood
Bone marrow
Purified cells from the above tissues

Stem Cells Detected

In vitro, hematopoietic multipotential stem cell (CFC-GEMM 1)
In vitro, lympho-hematopoietic primitive stem cell (HPP-SP 2)

Click image for larger view

Examples of Measuring Stem Cell "Quality" Using HALO®-96 SPC-QC

Normally, the CFC-GEMM 1 stem cell population would suffice for routine assessment. However, a stem cell product may contain different proportions of stem cell populations. A product with few, if any, primitive stem cells will only result in short-term engraftment and reconstitution. A product with few multipotential stem cells may be misinterpreted as containing few, if any, stem cells, even if sufficient primitive stem cells were present, but not detected. These and other possibilities could occur during the processing procedure if the latter had not been properly evaluated or may occur during the cryopreservation or cell thawing process, if these had not been sufficiently evaluated. The results shown in the upper graph demonstrate that umbilical cord blood mononuclear cells exhibit the lowest proliferation ability compared to bone marrow and mobilized peripheral blood (mPB) CD34⁺ cells for both the primitive and more mature stem cell populations. Please note that mPB cells are shown at 1,000 CD34⁺ cells/well, while bone marrow and cord blood are shown at 5,000 MNC/well.

The lower graph illustrates a number of important aspects of the HALO® Platform and HALO®-96 SPC-QC, in particular. HALO®-96 SPC-QC can be setup so that results can be obtained on day 5, 6 or 7. If greater assay sensitivity is required, the cultures can be processed to measure intracellular ATP on day 6 or 7, instead of day 5. For stem cells, there is a 3 fold increase in proliferation ability within 2 days when measured on days 5 and 7. This is because the cells are proliferating exponentially. However, after day 5, differentiation begins to increase. HemoGenix® recommends completing the assay on day 5, but if scheduling is a problem, it can be extended to day 7. Please note, however, that once day 5, 6 or 7 has been elected as the day of processing, ALL future assays should be processed on the same day, otherwise it will not be possible to properly compare results between assays.

The exponential increase in proliferation seen between days 5 and 7, and which continues to increase until about day 10, can only be detected using a proliferation assay such as HALO®. The CFU/CFC assay will not provide this information, since once a colony is formed, it will only get larger. The size of the colony is an indication of the primitiveness of the cells forming that colony.This information is very difficult to quantify and is lost to the user. The number of colonies counted between days 7 and 10 remains the same. Please refer to the graph shown on the HALO®-96 PCA^EQ page.

HALO®-96 SPC-QC and HALO®-96 PCA^EQ

Please note that although both these assays incorporate SEC and bioluminomics™ technology, they do not measure the same cell populations and therefore one cannot be used in place of the other.

Use

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

Plate luminometer or multiparameter plate reader with "glow" luminescence measuring capability.

Kit Contents

HALO®-96 SPC-QC Master Mix for HPP-SP and/or CFC-GEMM and/or background control
ATP standard
ATP high and low controls
ATP Enumeration Reagent
Sterile, 96-well plates
Non-sterile, 96-well plates
Sterile, adhesive foil covers
Instruction manual

HALO®-96 SPC-QC Assay Kit Manual

To download the HALO®-96 SPC-QC manual, please go to the Downloads Page.

Related Assays

HALO®-96 PQR

A Lympho-Hematopoietic Stem Cell Potency, Quality and Release Assay prior to Transplantation

Buy HALO®-96 PQR assay kits

Available as assay kits and
contract research services.

Stem Cell Potency

Stem cell potency is the quantitative measurement of stem cell proliferation potential.
Stem cell potency predicts engraftment potential, but not "time to engraftment", which is the ability of stem cells to reconstitute the hematopoietic system by differentiating and maturing into a specific number of neutrophils, platelets and erythrocytes. (To determine time to engraftment, use HALO®-96 PMT).

HALO®-96 PQR Applications

To determine stem cell potency, quality and acceptance limits for release criteria in a single assay.

NOTE: Although the community norm is to use the colony-forming unit (CFU) assay, such as CAMEO™-4, an equivalent and alternative assay, such as HALO®-96 PQR is acceptable by the FDA, since it satisfies "the requirements of the applicable statutes and regulations". In addition, AABB Standards for Cellular Therapy Services also allows "equivalent functional assays for any product", and the National Marrow Donor Program (NMDP) allows testing "consistent with AABB Standards".

Advantages of using HALO®-96 PQR prior to Stem Cell Transplantation

HALO®-96 PQR is the only commercially available and patented, reference standard-based stem cell potency assay for mobilized peripheral blood, umbilical cord blood or bone marrow that complies with FDA regulations for potency assays.
HALO®-96 PQR uses a 3-step procedure to reliably estimate the stem cell potency ratio and "quality" of stem cell populations.
Stem cell potency ratio and "quality" together help define whether a stem cell product can be used for transplantation.
Requires as few as 90,000 cells to perform the assay.
Instrument-based, incorporating an external standard and controls for assay standardization and validation according to FDA regulations as well as a reference standard of the material being measured.
Allows for intra- and inter-laboratory comparisons.
Assay can be completed on day 5, 6 or 7 of culture.
An FDA and AABB alternative potency assay.
Incorporates Suspension Expansion Culture (SEC) and Bioluminomics™ Technology, the most advanced and cost-effective lympho-hematopoietic in vitro assay system available.