HALO® SC-IPS 

HALO® RS

An Assay to Determine the Identity, Purity and Strength (Potency) for a Stem Cell Sample from Umbilical Cord Blood, Bone Marrow and Mobilized Peripheral Blood Compared to a Reference Standard
 

Buy HALO® SC-IPS Assay Kits
 

Buy HALO® RS Assay Kits


A complete bioluminescence assay to determine stem cell identity, purity and strength (potency) for umbilical cord blood, bone marrow and mobilized peripheral blood.

 

  • SC = Stem Cell
  • IPS = Identity, Purity, Strength. These are the parameters that the U.S. Food and Drug Administration (FDA) requires for traditional drugs and now biological drugs, such as umbilical cord blood and products for regenerative medicine.
  • Identity defines the "active ingredients" that are responsible for the intended effect or response. For a hematopoietic cell therapy products, only the stem cells are the "active" ingredients and must therefore be identified.
  • Purity defines whether the "active" ingredients are free of impurities and whether these impurities influence the intended effect or response. For hematopoietic cell therapy products, impurities can dilute and mask the identity of the "active" ingredients thereby leading to serious underestimation of the stem cell response. 
  • Strength is considered by the FDA to be equivalent to potency, which predicts the intended effect or response. The strength or potency of the "active" stem cell ingredients predicts stem cell engraftment prior to transplantation and therefore provide the measurement assurance to release the product for use in a patient. 
  • To determine stem cell identity, purity and strength in a single assay for umbilical cord blood, bone marrow or mobilized peripheral blood or purified cells to be used for transplantation into 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 compliant potency assay for hematopoietic cell therapy products.
  • HALO® SC-IPS has a Master File Number with the FDA that can be referenced in an IND or BLA to the FDA. For more information, please contact Preferred Cell Systems™.
  • HALO® SC-IPS methodology allows different stem cell populations to be distinguished from progenitor cell populations, thereby identifying the "active" stem cell ingredients. 
  • The results identifying the stem cells also indicates the purity of the stem cell fraction being determined.
  • The combined analysis of identity and purity allows the strength of the stem cell product to be determined in a single assay.
  • Reference standards for umbilical cord blood, bone marrow, mobilized peripheral blood and purified cells are also available to establish in-house reference standards using HALO® RS and to determine the potency ratio (measurement of potency) using HALO® SC-IPS.
  • Measurement of strength or potency is performed using a modified "Slope-Ratio Concentration-Response Model" standardized by the U.S. Pharmacopeia.
  • Not 1, but 2, primitive stem cell populations are measured using HALO® SC-IPS. This is because strength or potency using a single cell population is too inaccurate. 
  • Requires as few as 90,000 cells to perform the assay from a sample of about 0.1mL.
  • HALO® SC-IPS is instrument-based, with calibration and standardization capability that can be compared to known measurement assurance parameters to ensure that the assay is working correctly and the results will be trustworthy.
  • Assay standardization allows intra- and inter-laboratory comparisons.
  • HALO® SC-IPS and HALO® RS are completed in 7 days to provide the highest sensitivity.
  • Both HALO® SC-IPS and HALO® RS are available with serum-free hematopoietic stem cell growth media upon request.

Principle of ATP Bioluminescence Cell Proliferation Assays

All ATP bioluminescence assays kits from Preferred Cell Systems™ include an ATP standard(s) and controls.  Performing the ATP standard curve and controls is highly recommended. The controls are used to calibrate the instrument and the ATP standard is used to standardize the assay. The values obtained provide the measurement assurance parameters that allow you, the user, to ensure that the results obtained can be trusted.

An example of an ATP standard curve and control results are shown in the diagram below and actual values for the kit being used are given in the Technical Manual. If the values obtained are within the ranges given in the Technical Manual, the user can process the samples knowing that the results will be trustworthy.

Calibration and standardization of an ATP bioluminescence assay from Preferred Cell Systems™ - Measurement Assurance
 
  • 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® SC-IPS is used. HALO® SC-IPS does not include RS material. You will use your in-house established RS to measure the potency ratio of the sample. 
  • In vitro, hematopoietic multipotential stem cell (SC-GEMM 1)
  • In vitro, lympho-hematopoietic primitive stem cell (SC-HPP 2)

When ordering HALO®, please use the Catalog Number and Species. You can quickly find these products by copying the catalog number and pasting it in the advanced search.
Advanced Search

HALO® RS Assays Kits Available to Establish
an In-House Reference Standard (RSa)
Populations Measured(*) Catalog Number Cell Type(#) Number of Plates/Kit Format
SC-GEMM 1
SC-HPP 2
K2-PRS-1CB   Cord blood 1 Low serum
SC-GEMM 1
SC-HPP 2
K2-PRS-1CB34 Cord Blood CD34 1 Low serum
SC-GEMM 1
SC-HPP 2
K2-PRS-1mPB   Mobilized peripheral blood 1 Low serum
SC-GEMM 1
SC-HPP 2
K2-PRS-1mPB34   Mobilized peripheral blood CD34 1 Low serum
SC-GEMM 1
SC-HPP 2
K2-PRS-1BM   Bone marrow 1 Low serum
SC-GEMM 1
SC-HPP 2
K2-PRS-1BM34  Bone Marrow CD34 1 Low serum
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1CB   Cord blood 1 Serum-Free
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1CB34 Cord Blood CD34 1 Serum-Free
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1mPB   Mobilized peripheral blood 1 Serum-Free
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1mPB34   Mobilized peripheral blood CD34 1 Serum-Free
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1BM   Bone marrow 1 Serum-Free
SC-GEMM 1
SC-HPP 2
K2SF-PRS-1BM34  Bone Marrow CD34 1 Serum-Free

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

(#): 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 Preferred Cell Systems™ for more information.

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

 

HALO® SC-IPS Assays Kits Available to Determine
Identity, Purity and Strength (Potency)
Populations Measured (*) Catalog Number Cell Type Number of Plates/Kit Format
SC-GEMM 1
SC-HPP 2
K2-PQR-1 Any hematopoietic tissue(1,2) 1 Low serum
SC-GEMM 1
SC-HPP 2
K2SF-PQR-2 Any hematopoietic tissue(1,2) 1 Serum-Free


(*): 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): A RS should 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.

Bulk assays are available upon request. Please contact Preferred Cell Systems™

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

The recommended 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.

This section is in the process of being updated. Please return soon.

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

 

Reems JA, Hall KM, Gebru LH, Taber G, Rich IN. Development of a Novel Assay to Evaluate the Functional Potential of Umbilical Cord Blood Progenitors. Transfusion. 48:620-628 (2008).

Rich IN. Potency, Proliferation and Engraftment Potential of Stem Cell Therapeutics: The Relationship between Potency and Clinical Outcome for Hematopoietic Stem Cell Products. J Cell Sci Therapy. (2013).

Patterson J, Moore CH, Palser E, Hearn JC, Dumitru D, Harper HA, Rich IN. Detecting Primitive Hematopoietic Stem Cells in Total Nucleated and Mononuclear Cell Fractions from Umbilical Cord Blood Segments and Units. J Translational Medicine 13:94 (2015).

Rich IN. Improving Quality and Potency Testing for Umbilical Cord Blood: A New Perspective.
Stem Cells Translational Medicine. 4:967-973 (2015).

Hall, KM, Harper H, Rich IN. Hematopoietic Stem Cell Potency for Cellular Therapeutic Transplantation. In: Advances in Hematopoietic Stem Cell Research, R. Pelayo (Ed). In Tech. ISBN:978-953-307-930-1 (2012).

Harper H and Rich IN. Measuring the Potency of a Stem Cell Therapeutic. In: Stem Cell Protocols. Methods in Molecular Biology, Vol. 1235,  Rich IN (Ed). Published by Humana Press (2015).

Harper H and Rich IN. Bioluminescence Potency Measurement of Cellular Therapy Products. In: Cellular Therapy: Principles, Methods, and Regulations, 2nd Edition, Eds. Areman EM and Loper K. Published by AABB.

 

For Research Use Only. Not for clinical diagnostic use.

Luminescence or multimode plate reader capable of measuring "glow" luminescence.

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

HALO® SC-IPS Kit Contents

  • HALO® SC-IPS 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

HALO® SC-IPS QuickGuides

A complete bioluminescence assay to determine stem cell identity, purity and strength (potency) for umbilical cord blood, bone marrow and mobilized peripheral blood.


A complete bioluminescence assay to to establish reference standards for HALO® SC-IPS for umbilical cord blood, bone marrow and mobilized peripheral blood.