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Products - Stem Predict™/ HALO®     Page   1  |  2  |  3  
 
  • StemPredict

    3-Day Assays

  • HALO® Quality

    Control Assays

  • HALO® Potency

    HALO® RS Assays

  • HALO® TE: Time

    to Engraftment Assays

  • HALO® PMT: Patient

    Monitoring Assays

STEMpredict™

STEMpredict™ HT

A complete, 3-day culture assay kit for hematopoietic stem cell "quality", viability and functionality to assess and triage cord blood unit bankability and optimize mobilized peripheral blood collection with high-throughput capability

 

GO TO Preferred Cell Systems

 

 

Uses/Applications of STEMpredict™
  • A specialized 3-day ATP bioluminescence HALO® assay performed on fresh umbilical cord blood prior to cryopreservation.
  • STEMpredict™ is used to ensure that the cord blood stem cells are viable and have sufficient functionality to be permanently stored.
  • STEMpredict™ HT is the only assay that can be used for high-throughput umbilical cord blood screening using either 96- or 384-well plate formats.
  • STEMpredict™ is also for mobilized peripheral blood, collected by apheresis, to determine if stem cells have been sufficiently mobilized.

 

Benefits of Using STEMpredict™ in the Stem Cell Processing Laboratory
  • STEMpredict™ is the only assay that can be used for high-throughput, cord blood screening using either 96- or 384-well plate formats.
  • Optimizes mobilized peripheral blood collection and use.
  • Predicts stem cell growth, functionality and viability in 3 days.
  • Replaces all other functional assays with non-subjective, quantitative data.
  • Fast, easy to learn and use, and fully standardized allowing sample results to be directly compared with each other.
  • Incorporates ATP bioluminescence  technology, the most advanced and sensitive assay readout system available.
  • Incorporates proprietary Suspension Expansion Culture™ (SEC™) growth medium technology for optimal cell culture, growth and assay sensitivity.
  • Results within 30 min after cell culture.
  • Always reliable and reproducible, with low coefficients of variation (CVs).
  • FDA, AABB or FACT alternative “quality” assay.
  • STEMpredict@ is a Best Practices Assay and is one of the cell therapy assays included in the HemoGenix® Proficiency Testing Program using HALO®-PT

 Principle, Sensitivity and Parameters of ATP Bioluminescence Assays

Download information on the principle of ATP bioluminescence assays, sensitivity and other parameters

 STEMpredict™ Assays: Availability

 

Please note that STEMpredict™ is not a potency assay. To measure hematopoietic stem cell potency, please see HALO®-Potency.

  

STEMpredict™ and STEMpredict™ HT Assays Available

Assay Type Plate Format

Catalog Number

(Low serum / Serum-free)

Number of Plates/Kit

Number of Samples/Kit

STEMpredict 96-well plate

K2-SP-1 / K2-SPSF-1

1

8

STEMpredict 96-well plate

K2-SP-2 / K2-SPSF-2

2

16

STEMpredict 96-well plate

K2-SP-4 / K2-SPSF-4

4

32

STEMpredict HT

384-well plate

K3-SP-4 / K3-SPSF-1

1

32

STEMpredict HT

384-well plate

K3-SP-4 K3-SPSF-2

2

64

STEMpredict HT

384-well plate

K3-SP-4 / K#-SPSF-4

4

128

To perform STEMpredict™, high throughput, a liquid handler (robot) is required 

 Tissues used with STEMpredict™
  • Umbilical cord blood
  • Mobilized peripheral blood
  • Bone marrow
  • Purified cells from any of the above

 Cell Purity Requirements for Detection
Cell Purity Requirements for Detection

The recomended cell purity is a mononuclear cell (MNC) fraction or higher purity. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that dilute, mask and severely underestimate and even inhibit the detection of rare primitive stem cells.

 Examples

 

 

STEM predict: How STEMpredict predicts whether a cord blood unit has high quality cord blood stem cells

Click image for larger view


STEMpredict™ for Cord Blood Stem Cells

The graph shows an example of how STEMpredict™ predicts stem cell growth, functionality and viability for umbilical cord blood cells. If cells are metabolically viable (which is not the same as being viable by dye exclusion methods), they will demonstrate cellular and mitochondrial integrity by producing intracellular ATP (iATP). The iATP concentration, in turn, directly correlates with proliferation ability. If a quantitative difference in proliferation ability between stimulated and unstimulated (background) stem cells exists, as shown in the graph, the stem cells from the product are predicted to proliferate and grow. In this way, STEMpredict™ not only detects whether the cells are metabolically viable, but also whether they will exhibit cellular functionality and proliferate. The assay therefore measures three parameters simultaneously.


This same concept and principle is applied to mobilized peripheral blood, bone marrow and purified hematopoietic cell products. It is also the cornerstone of all HemoGenix® cellular therapeutic assay kit products, including HALO® PCAEQ, HALO® SPC-QC, HALO® TE, HALO® PMT, MSCGlo™ HuQC and MSCGlo™ PQR. 

 Use and Major Equipment Required

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

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

 Assay Kit Contents
  • STEMpredict™ stem cell and control Master Mix
  • ATP standard
  • ATP high and low controls
  • ATP Enumeration Reagent
  • Sterile, 96- or 384-well plates for cell culture
  • Non-sterile 96-well plates (for assay calibration and standardization)
  • Sterile, adhesive foil covers to maintain sterility in unused wells
  • Instruction manual

 STEMpredict™ Information Flyer, QuickGuide and Assay Manual

 Related Assays

HALO® QC

A 5-day, complete assay kit to optimize and measure the health and quality of cord blood, bone marrow and mobilized peripheral blood stem cell

 

GO TO Preferred Cell Systems

 

 

 
Stem Cell "Quality" and Stem Cell Potency
  • Stem cell "quality" is defined as the proliferation ability of stem cells at a specific point in time and at a particular cell dose.
  • Stem cell "quality" cannot be measured using minimal testing criteria, (e.g total nucleated cell (TNC) count, viability, viable CD34 or a CFU assay), because these do not quantitatively measure the proliferation ability of stem cells.
  • HALO® QC is NOT a potency assay. To measure stem cell potency, please see HALO® Potency).
 
Uses of HALO® QC
  • To quantify the proliferation ability or status ("quality") of stem cells in a stem cell product. Please note that this is not the same as stem cell potency.
  • To compare stem cell "quality" before and after a specific proecdure (e.g. red blood cell depletion of cord blood).
  • To determine stem cell "quality" pre- and post cryopreservation.
  • To determine the effect on stem cell "quality" of equipment and bags used for stem cells.
  • To determine stem cell processing stability.
  • Gene targeting.
  • Culture and expansion of stem cells in vitro and in bioreactors.
  • To promote "Best Practice Criteria Testing".
 
Benefits of using HALO® QC in the Stem Cell Processing Laboratory
  • HALO® QC quantitatively measures viability and proliferation ability and cellular functionality that cannot be enumerated using total nucleated counts (TNC), viability or flow cytometric detection of membrane expression markers.
  • HALO® QC determines the "quality" of mature, hematopoietic stem cells (SC-GEMM) alone or SC-GEMM and more primitive lympho-hematopoietic stem cells (SC-HPP).
  • Instrument-based, incorporating an external standard and controls for assay calibration and standardization.
  • Has been validated according to FDA bioanalytical method validation guidelines and can be validated in-house.
  • Compare results over time for intra- and inter-laboratory comparison studies.
  • Assay can be completed in just 5 days or can be extended to 7 days with a 3-fold increase in sensitivity (see below).
  • Faster, easier to use and more accurate than any traditional CFU assay, even using automated colony counting.
  • HALO® QC is a CFU-substitute proliferation assay allowed by the FDA, AABB and NetCord-FACT. Results from HALO® QC can be uploaded to the NMDP cord blood database.
  • HALO® QC incorporates proprietary Suspension Expansion Culture™ (SEC™) Growth Medium and ATP bioluminescence technology, the most advanced and cost-effective lympho-hematopoietic in vitro assay system available for the stem cell processing laboratory.

 Principle, Sensitivity and Parameters of ATP Bioluminescence Assays

Download information on the principle of ATP bioluminescence assays, sensitivity and other parameters

 Stem Cells Detected
  • In vitro, hematopoietic multipotential stem cell (SC-GEMM 1)
  • In vitro, lympho-hematopoietic primitive stem cell (SC-HPP)

 

HALO® QC Assays Available

Assay Type Cell Type Catalog Number Number of Plates/Kit

Number of Samples/Kit

Click to see price

HALO®

SPC-QC

SC-GEMM 1

K2-1QC-1

1

16

Price
HALO®
SPC-QC
SC-GEMM 1

K2-1QC-2

2

32

Price
HALO®
SPC-QC
SC-GEMM 1

K2-1QC-4

4

64

Price
HALO®
SPC-QC
SC-GEMM 1 + SC-HPP 2

K2-2QC-2

2

16

Price
HALO®
SPC-QC
SC-GEMM 1 + SC-HPP 2

K2-2QC-4

4

32

Price

 Tissues

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

 Cell Purity Requirements for Detection

The recomended cell purity is a mononuclear cell (MNC) fraction or higher purity. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that dilute, mask and severely underestimate and even inhibit the detection of rare primitive stem cells.

 Examples

HALO-96 SPCQC: Demonstrating stem cell quality in cord blood, bone marrow and mobilized peripheral blood

Click image for larger view

 

HALO-96 SPCQC: Demonstrating how culture time can influence assay sensitivity of different cell populations

Click image for larger view

Examples of Measuring Stem Cell "Quality" Using HALO® QC

Normally, the SC-GEMM 1 (methylcellulose, CFC-GEMM 1 equivalent) 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® QC, in particular. HALO® 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® PCA page.

 

HALO® QC and HALO® PCA

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

 Use and Major Equipment Required

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

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

 Assay Kit Contents
  • HALO® QC Master Mix for SC-HPP and/or SC-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® QC Information Flyer, QuickGuide and Assay Manual

 Related Assays

HALO® Potency and  

HALO® Reference Stanadard (RS) Assays

A 7-Day, FDA-compliant and validated, bioluminescence complete stem cell potency, quality and release assay for umbilical cord blood, bone marrow and mobilized peripheral blood 

 

GO TO Preferred Cell Systems

 

 

Articles on stem cell potency using HALO-Potency

Click image for larger view

What is Hematopoietic Stem Cell Potency?
  • Stem cell potency is the quantitative and validated measurement of proliferation potential or capacity (biological activity) of the "active" stem cell components of a cord blood, mobilized peripheral blood or bone marrow product, which, when administered to the patient, produces the intended response of engraftment.
  • Stem cell proliferation potential or capacity correlates directly with stem cell self-renewal and primitiveness or "stemness". Thus, the greater the proliferation potential, the greater the self-renewal capacity and the more primitive the stem cell population. The more primitive a stem cell population, the greater its potency.
  • Potency must correlate with a response, but not necessarily the clinical outcome. Thus, stem cell potency does not predict or correlate with "time to engraftment", which is the ability of the engrafted stem cells to produce short-term reconstitution by differentiating and maturing into a specific number of neutrophils, platelets and erythrocytes. (To determine time to engraftment, please use HALO® PMT).

 

Uses of HALO® Potency / HALO® RS
  • To determine stem cell potency and "quality" in a single assay on hematopoietic cell therapy products prior to transplantation into a patient.
  • To use both stem cell potency and "quality" to define release criteria and determine if a product can be released for use in a patient.
  • To establish in-house reference standards (using HALO® RS) that allow the potency ratio (measure of potency) to be determined.
  • To use an FDA complient potency assay for hematopoietic cell therapy products.
  • To promote "Best Practice Criteria Testing". 

 

Benefits of using HALO® Potency Prior to Stem Cell Transplantation
  • HALO® Potency has been assigned an FDA Master File Number that can be referenced in an IND or BLA to the FDA. For more information, please contact HemoGenix®.
  • HALO® Potency is the only commercially available, 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® Potency incorporates the "Slope-Ratio Concentration-Response Model" standardized by the U.S. Pharmacopeia.
  • HALO® Potency reliably estimates the stem cell potency ratio and "quality" of two stem cell populations simultaneously against a reference standard to predict whether a cord blood, mobilized peripheral blood or bone marrow unit can be transplanted into a patient.
  • Requires as few as 90,000 cells to perform the assay.
  • Instrument-based, incorporating an external standard and controls for assay calibration, standardization and validation according to FDA regulations.
  • Allows for intra- and inter-laboratory comparisons.
  • HALO® Potency and HALO® RS are completed in 7 days to provide the highest sensitivity.
  • HALO® Potency is allowed by the FDA, AABB and NetCord-FACT as alternative validated potency assays.
  • Incorporates proprietary Suspension Expansion Culture™ (SEC™) stem cell growth media and ATP bioluminescence technology, the most advanced and cost-effective lympho-hematopoietic in vitro assay system available.
  • HALO® Potency / HALO® RS are Best Practices Assays and cell therapy assays included in the Hemogenix® Proficiency Testing Program using HALO®-PT.

 Principle, Sensitivity and Parameters of ATP Bioluminescence Assays

Download information on the principle of ATP bioluminescence assays, sensitivity and other parameters

 Difference between HALO® Potency and HALO® RS
  • To measure potency, a reference standard (RS) of the same material as the sample is required so that the potency ratio can be determined. The inclusion of a RS to measure potency is an FDA requirement. Universal or global reference standards for cell therapy products are not available. As a result, it is necessary for individual centers to establish their own RS. To accomplish this, a standardized and validated assay platform is required.
  • HALO® RS is used to establish your first in-house RS for a specific tissue. The assay includes a vial of cord blood, peripheral blood or bone marrow cryopreserved cells that are used to compare to your own preparation. HALO® RS is only required once to set up your first RS cell preparation. All further reference standards establish in-house will be compared to the previous RS. (See below).
  • To measure potency of your samples prior to use, HALO® Potency is used. HALO® Potency does not include RS material. You will use your in-house established RS to measure the potency ratio of the sample. 

 Stem Cell Populations Detected (The "Active" Components Measured in a Potency Assay
  • In vitro, hematopoietic multipotential stem cell (CFC-GEMM 1)
  • In vitro, lympho-hematopoietic primitive stem cell (HPP-SP 2)

 HALO® RS and HALO® Potency Assays Available

HALO® RS Assays Kits Available to Establish an

In-House Reference Standard (RS)

Assay Type Cell Type(#) Populations Measured(*) Catalog Number Number of Plates/Kit

Number of Samples/Kit

Click to see price

HALO® RS(a)

Cord blood SC-GEMM 1 + SC-HPP 2

K2-PRS-1CB

1

1 + RS

Price
HALO® RS Cord Blood SC-GEMM 1 + SC-HPP 2

K2-PRS-2CB

2

3 + RS

Price
HALO® RS Mobilized peripheral blood SC-GEMM 1 + SC-HPP 2

K2-PRS-1mPB

1

1 + RS

Price

HALO® RS

Mobilized peripheral blood SC-GEMM 1 + SC-HPP 2

K2-PRS-2mPB

2

3 + RS

Price
HALO® RS Bone marrow SC-GEMM 1 + SC-HPP 2

K2-PRS-1BM

1

1 + RS

Price
HALO® RS Mone Marrow SC-GEMM 1 + SC-HPP 2

K2-PRS-2BM

2

3 + RS

Price

 

(#): HALO® RS assay kits are also available to help establish reference standards for purified CD34+ cells from each of the 3 tissues. Please go to the online catalog or contact HemoGenix for more information.

(*): A minimum of 2 stem cell populations must be measured. Potency cannot be determined using a single cell population. 

(a): Prior to measuring potency, a reference standard (RS) must be established to which the sample can be compared and the potency ratio determined. 

 

HALO® Potency Assays Kits Available to Determine

Stem Cell Potency, Quality and Release

Assay Type Cell Type Populations Measured (*) Catalog Number Number of Plates/Kit

Number of Samples/Kit

Click to see price

HALO® Potency

Any hematopoietic tissue(1) SC-GEMM 1 + SC-HPP 2

K2-PQR-1

1

1 + RS

Price
HALO® Potency Any hematopoietic tissue(1) SC-GEMM 1 + SC-HPP 2

K2-PQR-2

2

2 + 2 RS(2) or

3 + 1 RS(2)

Price
HALO® Potency Any hematopoietic tissue(1) SC-GEMM 1 + SC-HPP 2

K2-PQR-4

4

6 + 1 RS(2) or 

4 + 4 RS(2)

Price

(*): A minimum of 2 stem cell populations must be measured. Potency cannot be determined using a single cell population.

(1): Includes mononuclear cells (MNC) from umbilical cord blood, mobilized peripheral blood, bone marrow or purified cells (e.g. CD34+ or CD133+) from these tissues.

(2): One RS must always be performed when an unknown sample is measured. If more than 1 sample is to be measured, a single RS of the same cell type can be used to compare with all samples performed on the same day.

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

 Cell Purity Requirements for Detection

The recomended cell purity is a mononuclear cell (MNC) fraction or higher purity. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that dilute, mask and severely underestimate and even inhibit the detection of rare primitive stem cells.

 Examples

HALO-Potency: How Stem Cell Potency, Quality and Release Criteria are Determined for mobilized peripheral blood

Click image for larger view

 

HALO-Potency: How Stem Cell Potency, Quality and Release Criteria are Determined Bone Marrow

Click image for larger view

 

HALO-Potency: How Stem Cell Potency, Quality and Release Criteria are Determined Umbilical Cord Blood

Click image for larger view

 

HALO-Potency: How both stem cell potency and quality are determined for 28 samples of umbilical cord blood

Click image for larger view

 

 

 

How Stem Cell Potency, Quality and Release Criteria are Determined using HALO® Potency

For background information on potency, please refer to Background: Cellular Therapy. In addition, the basic methodology for measuring stem cell potency was published in 2016 in Cellular Therapy: Principles, Methods and Regulations (Eds. Areman EM, Loper K) and entitled "Bioluminescence Potency Measurement of Cellular Therapy Products".

 

Download the article: Karen M. Hall, Holli Harper and Ivan N. Rich (2012). Hematopoietic Stem Cell Potency for Cellular Therapeutic Transplantation, Advances in Hematopoietic Stem Cell Research, Rosana Pelayo (Ed.), ISBN: 978-953-307-930-1, InTech.

Download the review article: I.N. Rich (2013). Potency, Proliferation and Engraftment Potential of Stem Cell Therapeutics: The Relationship vetween Potency and Clinical Outcome for Hematopoietic Stem Cell Products. J. Cell Science & Therapy. This is an Open Access article.

 

To measure potency, or more specifically, the potency ratio, three basic requirements need to be met. These are:

  1. A standardized and validated assay readout.
  2. The ability to measure the "active stem cell ingredients", in a dose-dependent manner. Please note that a total nucleated cell (TNC) fraction should not be used to measure potency, since it contain high concentrations of cell impurities that dilute and mask the stem cells. See a comparison between the TNC and MNC fraction. Click here.
  3. A reference standard of the same material

All three of these requisites, as well as other regulatory requirements, have been incorporated into HALO® Potency. The "active ingredients" are the hematopoietic stem cells. From the discussion on lympho-hematopoiesis in Background: Lympho-Hematopoiesis and other areas of this website, it will be appreciated that assays are not available to detect every stem cell stage in the stem cell compartment continuum. For this reason, two distinct stem cell populations are presently detected using HALO® Potency, which together provide a good estimation of stem cell potency in a sample or product.
A stem cell dose response is required to measure proliferation potential. The later is defined as the slope of the linear regression curve fit of the dose response for each stem cell population measured. The steeper the slope of the stem cell dose response, the greater the proliferation potential and the more primitive the stem cell population. The greater the proliferation potential, the greater the potency. Therefore, the potency ratio is the ratio of the slope of the stem cell population of the sample compared to that of the reference standard. Please note that when performing potency measurements, it is necessary to establish primary, secondary and even tertiary in-house reference standards. The reference standard included in the HALO® Potency assay kit is meant as a "starter" for in-house reference standards. 

The results shown in the three graph panels demonstrate how potency ratios are calculated and the two parameters necessary for defining the release criteria of a product. The potency of the reference standard is always 1. The acceptance / rejection level for stem cell "quality" is usually provided from historical data. However, intracellular ATP (iATP) concentrations less than 0.03-0.04µM is an indication that cells cannot sustain proliferation or may be dead. Stem cell "quality" measurements greater than the arbitrary acceptance level and potency ratios usually greater than 1 for the stem cell populations measured would provide the premise for accepting a product.The last panel shows stem cell "quality" and potency ratios for 28 cryopreserved cord blood clinical samples.


For more information on potency testing of hematopoietic stem cell products, please download the HALO® Potency Information Flyer and the White Paper on potency testing.

 Use and Major Equipment Required

For Research Use Only. Not for clinical diagnostic use

Major Equipment Required

A plate luminometer or multiparameter plate reader capable of measuring "glow" luminescence is needed for this assay.

 Assay Kit Contents
HALO®-RS Kit Contents:
  • Vial of cryopreserved reference standard umbilical cord blood, mobilized peripheral blood, bone marrow mononuclear cells (MNC) or purified cells (CD34+) from the respective tissue
  • HALO® Master Mixes for both SC-HPP 2 and SC-GEMM 1 stem cell populations
  • ATP standards
  • ATP controls
  • ATP Enumeration Reagent
  • Sterile, 96-well plate(s)
  • Non-sterile, 96-well plate(s)
  • Sterile, adhesive foil covers
  • Assay manual

HALO®-Potency Kit Contents
  • HALO® Potency Master Mixes for SC-HPP 2 and SC-GEMM 1 stem cell populations
  • ATP standard
  • ATP controls
  • ATP Enumeration Reagent
  • Sterile, 96-well plates
  • Non-sterile, 96-well plates
  • Sterile, adhesive foil covers
  • Assay manual
 HALO® Potency and HALO®-RS Information Flyer, QuickQuides and Assay Manual

HALO® Potency and RS Information Flyer

HALO® Potency and RS QuickGuides coming soon

HALO® Potency and RS assay manual coming soon

 Frequently Asked Questions
Why do HALO® Potency and HALO® RS measure stem cells? Without stem cells, there would be no stem cell transplantation procedure. If stem cells are not detected and measured, the procedure is based on the false assumption that stem cells are present and exhibit high potency and quality. The high rate of graft failure has been attributed to low or lack of potency.Since stem cells are responsible for engraftment, these cells must be measured to assure high potency and quality.

Can TNC, viability, CD34 and CFU be used to measure potency? No. None of these tests either individually or together comply with regulations govening potency assays. In addition, potency must be measured using a dose response relationship and cannot be determined using a single point.

Both CFU and HALO® Potency are functional assays. Why isn't the CFU assay a potency assay? First it is not a validated assay, and a potency assay must be validated. Second, the CFU assay cannot accurately and reliably measure the "active ingredients" required for a potency assay. Third, the CFU assay does not incorporate standards and controls, which are a requirement for a potency assay. Fourth, a single point assay cannot be used for a potency determination; a dose response is required. Fifth, the CFU is a subjective assay that cannot and shouldnot be used for clinical use.

What is a Reference Standard for potency? Potency can only be determined when the sample is directly compared to a reference standard of the same material and purity. The potency of the reference standard is always 1 (one).

What is the measure of potency? The measure of potency is the potency ratio and is determined by a comparison of the sample with a reference standard of the same material and purity. A potency ratio >1 indicates that the sample can be used at a lower dose than the reference standard to produce the same response as the reference standard, and visa versa for a potency ration <1.

What is the "Slope-Ratio Concentration-Response Model"? The "Slope-Ratio Concentration-Response Model" is an alternative to the dose response statistical parallelism model used to measure the potency of traditional drugs. In the "Slope-Ratio Concentration-Response Model", the slope of the dose response curve for the sample is compare to that of the reference standard. This model is a U.S. Pharmocopeia standardized procedure. 

Can the ALDH assay be used as a potency or quality assay? No. One of the requirements for a potency assay is to measure the specific "active ingredients" or components, which in this case are the stem cells. The ALDH assay is not specific for stem cells; it is present in many other cell types and therefore cannot be used, by itself, as a potency assay. 

HALO® TE

A 5-day complete bioluminescence end point assay kit to determine time to engraftment of neutrophils, red blood cells and platelets after stem cell transplantaion

 

GO TO Preferred Cell Systems

 

 

Uses of HALO® TE
  • To rapidly predict and measure the time to neutrophil, erythroid and/or platelet engraftment after hematopoietic stem cell transplantation.
  • To promote "Best Practice Criteria Testing".

Benefits of using HALO® TE for Detecting Time to Engraftment
  • Measure the ability and amount of P-GM, P-BFU and/or P-Mk progenitor cell proliferation prior to detecting differentiated cells in a CFU assay.
  • Proliferation of progenitor cells can be detected within 5 days.
  • Allows for the detection of individual hematopoietic progenitor cell populations or all three simultaneously after transplantation.
  • Faster, easier and more accurate to use than any traditional CFU assay, even with automated colony counting.
  • Allows comparison of patient results over time due to inclusion of assay standards and controls.
  • HALO® TE is a validated assay.
  • Incorporates proprietary Suspension Expansion Culture (SEC) growth medium and ATP bioluminescence technology to produce the most advanced, accurate, sensitive and cost-effective assay system available.
  • HALO® TE is available with low serum or serum-free formulations of HALO® Master Mixes.
  • HALO® TE is a Best Practices Assay and one of the cell therapy assays included in the HemoGenix® Proficiency Testing Program using HALO® PT.

 Principle, Sensitivity and Parameters of ATP Bioluminescence Assays

Download information on the principle of ATP bioluminescence assays, sensitivity and other parameters

 Populations Detected

To determine Time to Engraftment:

  • P-BFU 1: Erythropoietic progenitor cell, used to predict red blood cell production.
  • P-GM 1: Granulocyte-macrophage progenitor cell, used to determine neutrophil engraftment.
  • P-Mk 1: Megkaryopoietic progenitor cell, used to predict platelet engraftment.

HALO® TE Assays Available

Assay Type Population Measured

Catalog Number

(Low serum /

Serum-Free)

Number of Plates/Kit

Number of Samples/Kit

HALO® TE

P-BFU 1

K2-BTE-1(1) /

K2-BTESF-1(2)

1

16(#)

HALO® TE

P-GM 1

K2-GMTE-1(1) /

K2-GMTESF-1(2)

1

16(#)

HALO® TE P-Mk 1

K2-MkTE-1(1) /

K2-MkTESF-1(2)

1

16(#)

HALO® TE P-BFU 1, + P-GM 1 + P-Mk 1

K2-3PTE-1(1) /

K2-3PTESF-1(2)

1

16(#)

(1): Low serum assays kits.

(2): Serum-free assay kits.

(#): These assay kits detect 16 samples for each cell population detected using 6 replicates/sample.

 Hematopoietic Stem Cell Product Transplantation Sources
  • Umbilical cord blood
  • Peripheral blood
  • Bone marrow biopsy

Tissues Used for Detection
  • Peripheral blood
  • Bone marrow biopsy

 Cell Purity Requirements for Detection

The recommended cell purity is a mononuclear cell (MNC) fraction. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that severely underestimate and even inhibit the detection of progenitor cells. 

 Use and Major Equipment Required

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

Luminescence plate reader or multiparameter plate reader with "glow" luminescence measuring capability.

 Assay Kit Contents
  • HALO® PT Master Mix(s) as either low serum or serum-free formulations
  • ATP standard
  • ATP high and low controls
  • ATP Enumeration Reagent
  • Sterile, 96-well plates
  • Non-sterile, 96-well plates
  • Sterile, adhesive foil covers
  • Assay manual
 Quick Guide and Assay Manual

Download the HALO® TE QuickGuide

Please navigate to the Resources section of the website to download the HALO®-96 TE Assay Manual

HALO® PMT

"Global" hematopoietic stem and progenitor cell complete assays for 5-, 6- or 7-populations to monitor patient reconstitution after transplantation.

 

GO TO Preferred Cell Systems

 

 

 

 

Uses of HALO® PMT
  • To detect and quantify "global" hematopoietic reconstitution.
  • To detect and quantify "global" lympho-hematopoietic reconstitution.
  • To promote "Best Practice Criteria Testing".

 

Benefits of using HALO® PMT for Patient Monitoring
  • Measurement of proliferation, rather than differentiation (CFU assays) allows prediction of cell growth providing faster results.
  • Distinguish between balanced and unbalanced cell lineage reconstitution.
  • Allows simultaneous detection of hematopoietic stem and progenitor cells to be measured at any time to demonstrate and follow reconstitution.
  • Faster, easier and more accurate to use than any traditional CFC assay, even with automated colony counting.
  • Results obtained in only 5 days.
  • Allows comparison of patient results with time due to inclusion of assay standards and controls.
  • Incorporate proprietary Suspension Expansion Culture (SEC) growth media and ATP bioluminescence technology, to produce the most advanced and cost-effective assay system available.

 Principle, Sensitivity and Parameters of ATP Bioluminescence Assays

Download information on the principle of ATP bioluminescence assays, sensitivity and other parameters

 Populations Detected

To determine "global" reconstitution:

  • 4-Population Assay for SC-GEMM 1, P-BFU 1, P-GM 1 and P-Mk 1
  • 5-Population Assay for SC-HPP 2, SC-GEMM 1, P-BFU 1, P-GM 1 and P-Mk 1
  • 7-Population Assay for SC-HPP 2, SC-GEMM 1, P-BFU 1, P-GM 1, P-Mk 1, P-Tcell, P-Bcell and background control

HALO® PMT Assays Available

Assay Type Population Measured

Catalog Number

(Low serum /

Serum-Free)

Number of Plates/Kit

Number of Samples/Kit

HALO® PMT

SC-GEMM, P-BFU, P-GM, P-Mk, Control(1)

K2-4PMT-5 /

K2-4PMTSF-5

5

16(#)

HALO® PMT

SC-HPP, SC-GEMM, 

P-BFU, P-GM, P-Mk, Control(2)

K2-5PMT-6 /

K2-5PMTSF-6

6

16(#)

HALO® PMT

SC-HPP, SC-GEMM,
P-BFU, P-GM, P-Mk,

P-Tcell, P-Bcell, Control(3)

K2-7PMT-8 /

K2-7PMTSF-8

8

16(#)

SC = Stem cell; P=progenitor cell

(1): To predict "global" hematopoietic reconstitution.

(2): To predict "global" hematopoietic reconstitution.

(3): To predict "global" lympho-hematopoietic reconstitution.

(#): These assay kits detect 16 samples for each cell population detected using 6 replicates/sample.

 Hematopoietic Stem Cell Product Transplantation Sources
  • Umbilical cord blood
  • Peripheral blood
  • Bone marrow biopsy

Tissues used for Detection
  • Peripheral blood
  • Bone marrow biopsy

 Cell Purity Requirements for Detection

The recomended cell purity is a mononuclear cell (MNC) fraction. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that severely underestimate and even inhibit the detection of progenitor cells. 

 Uses and Major Equipment Required

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

Luminescence plate reader or multiparameter plate reader with "glow" luminescence measuring capability.

 Assay Kit Contents
  • HALO® PMT Master Mix
  • ATP standard
  • ATP high and low controls
  • ATP Enumeration Reagent
  • Sterile, 96-well plates
  • Non-sterile, 96-well plates
  • Sterile, adhesive foil covers
  • Assay manual

 HALO® PMT Quick Guide and Assay Manual

Download the HALO® PMT Quick Guide

Please navigate to the Resources section to download the HALO®-96 PMT Assay Manual