HemoGenix^® offers two types of studies for assessing the quality and potency of potential clinical products. 

1. Complete Service, Full Report Study: Fully customized study that includes the Study Plan, Draft Text and Final Text Report with QA audit. This might include design and/or development of a new potency assay, its implementation and help with regulatory approval.
2. Rapid Potency Study: This would be for the routine assessment of potency of single or multiple samples either for clinical or verification purposes. The samples can be shipped to HemoGenix^® either individually or as multiple samples. The sample will usually be cryopreserved. Raw results will be provided in an Excel Workbook and a Certificate of Potency (CoPA, see below) will be provided for each sample. 

The type of study (1 or 2) will be determined by HemoGenix^®.

The following samples can be used for testing: 

1. Fresh samples of any volume down to about 0.1mL shipped cold, but not frozen.

2. Cryopreserved samples of any volume down to about 0.1mL shipped on dry ice or in a liquid nitrogen dewar.

3. Samples from umbilical cord blood, bone marrow, normal or mobilized peripheral blood or purified populations, e.g. CD34⁺, CD133⁺, used for hematopoietic cellular therapy.

4. Cultured or passaged mesenchymal stem/stromal cells (MSC) from any source.

5. ES, iPS or genetically modified cells. 

If samples contain cell impurities that might affect potency, such samples might require further processing to ensure that the best potency assessment can be performed.

The importance of measuring cell potency cannot be underestimated. It is probably THE most important parameter that must be measure prior to use of a clinical product. It is also one of the most misunderstood parameters and the assay can be the most complex.  

Potency is used to:

• Ensure consistency during manufacture of the product.
• Ensure stability of the product.
• Avoid product failure due to incorrect potency. This is particularly the case for umbilical cord blood, which demonstrates about 20% graft failure that has been attributed to lack of or low potency.
• Reduce potential toxicity.
• Predict whether the product can be released for use.

All of these reasons indicate that potency is a prospective measurement, not a retrospective measurement.

Any potency assay must include the following:

• A potency assay must be quantitative and validated assay.
• It must include measurement assurance parameters that allow the user to ensure not only that the assay is working correctly, but that the results are trustworthy.
• It must predict the intended biological response.
• It must include a reference standard of the same source of target cells processed to the same purity.
• The potency of the product should indicate the required dose to achieve the expected response. 

After all, the potency assay is meant to ensure that the clinical product is going to work as intended. 

HemoGenix^® has three potency assays readily available for Rapid Potency Studies. These include:

• HALO® SC-IPS for hematopoietic stem cell potency determinations for cryopreserved umbilical cord blood, bone marrow or mobilized peripheral blood samples.
• MSCGlo™ SC-IPS for cryopreserved MSC samples from virtually any source.
• STEMGlo™ SC-IPS for ES or iPS samples.     

"SC" denotes that these potency assays are for stem cells. "IPS" denotes that these assay measure stem cell Identity, Purity and Strength or Potency when compared directly to a reference standard of the same tissue source. 

Since all three potency assays measure (a) proliferation ability and, (b) proliferation potential, both stem cell "quality" and potency are determined in the same assay.

• Stem cell potency contract services are available for umbilical cord blood, mobilized peripheral blood, bone marrow and purified stem cells (e.g. CD34⁺) from these tissues.
• Stem cell potency and quality of the tissue sample is measured using HALO^® SC-IPS.
• Stem cell potency is determined by comparing a cell dose response with that of the reference standard.
• HALO^® SC-IPS is an FDA validated assay platform that complies with FDA and EMA guidelines and regulations for potency assays.
• HALO^® SC-IPS has a Master File with the FDA.
• Hematopoietic stem cell potency contract services are performed under GLP compliance.
• Hematopoietic stem cell potency of the sample is measured for the lympho-hematopoietic high proliferative potential - stem and progenitor (SC-HPP 2) cell and the primitive hematopoietic stem cell population (SC-GEMM 1) against a reference standard of the same tissue material.
• HemoGenix^® has established reference standards for umbilical cord blood, mobilized peripheral blood and bone marrow mononuclear cells that are used to determine the potency ratio of samples from the same tissues.
• Stem cell culture is performed using our proprietary Suspension Expansion Culture™ (SEC™) and standardized ATP bioluminescence readout technology, the most advanced and sensitive, non-subjective signal detection system available.
• Assay requires a minimum of only 120,000 sample cells.
• Results available usually after 7 days of culture to provide the greatest assay sensitivity.
• Thanks to our standardized ATP bioluminescence technology, results from different samples can be compared over time.
• All results are included in a Certificate of Potency Analysis (CoPA). The CoPA provides information on the stem cell potency ratio, stem cell €œquality€ and the release criteria. Final product release determination should be made by the Medical Director or similar authority and only after all information pertaining to the characterization of the product have been reviewed.

Please note that when considering stem cell potency of hematopoietic cellular therapeutic products, total nucleated cell count (TNC), dye exclusion viability and detection of viable CD34⁺ cells represent "minimum testing criteria" and are not potency assays. They do not comply with FDA or EMA guidelines or regulations. However, cell count and viability are necessary to use the cells to measure potency. The number of CD34⁺ cells also provides additional characterization information. The CFU assay is not a potency assay and is not compliant with potency assay regulations.  

MSCGlo™ SC-IPS and STEMGlo™ SC-IPS demonstrate many of the same benefits as HALO^® SC-IPS, since they were developed from the HALO^® assay.

• MSCGlo™ SC-IPS assays incorporate MSCs derived from different sources.
• STEMGlo™ SC-IPS assays use ES or iPS samples and reference standards provided by the Sponsor.
• Both assays determine "quality" and potency. Providing a reference standard is included, the identity and purity is also determined.
• Stem cell potency is determined by comparing a cell dose response with that of the reference standard.
• Both MSCGlo™ SC-IPS and STEMGlo™ SC-IPS are validated according to FDA and EMA guidelines and regulations for potency assays.
• Both MSC or ES/iPS potency contract services are performed under GLP compliance.
• HemoGenix^® has established reference standards for MSCs to determine the potency ratio. For ES and iPS cells, refernce standards should be provided by the Sponsor.
• Stem cell culture is performed using proprietary growth medium and standardized ATP bioluminescence readout technology, for most advanced and sensitive, non-subjective signal detection system available.
• Results available usually after 3-7 days of culture.
• Thanks to our standardized ATP bioluminescence technology, results from different samples can be compared over time.
• All results are included in a Certificate of Potency Analysis (CoPA). The CoPA provides information on the stem cell potency ratio, stem cell €œquality€ and the release criteria. Final product release determination should be made by the Medical Director or similar authority and only after all information pertaining to the characterization of the product have been reviewed.

Please note that when considering stem cell potency cell count and dye exclusion viability are not potency assays. However, cell count and viability are necessary to measure potency. Membrane markers used to identify MSCs, ES or iPS cells provide additional characterization information, but are not potency assays. The CFU-F assay used for MSCs is not a potency assay and is not compliant with potency assay regulations.  

Cell "quality" is simply defined as the ability of cells to proliferate using a single cell dose. Cell "quality" is often misrepresented as either sufficient cell numbers, high viability or sufficient cells exhibiting one or more other markers.

There are many applications where cell "quality" should be determined: These include, but are not limited to:

• Ascertaining the efficiency of new equipment and tools to fractionate and/or process cells.
• Optimizing cell culture protocols. 
• Optimizing a cryopreservation procedure.
• Assessing the "quality" of cryopreserved cells stored for long periods of time. 
• Optimizing a thawing procedure.
• Cell fractionation and separation procedures.
• Cell expansion procedures.
• Cell modification procedures. 

All of these rely on the fact that there is a "before" and "after" cell quality and in practice, any procedure or protocol should not affect the "quality" of cells. 

For this reason, HemoGenix^® developed standardized and validated assays to measure cell "quality" in the most used products for cellular therapy and regenerative medicine. These include:

• STEMpredict™ for triaging high from low "quality" hematopoietic stem cell product prior to cryopreservation.
• HALO^® QC for measuring hematopoietic stem cell quality.
• MSC™ QC for measuring mesenchymal stem cell quality.
• STEMGlo™ QC for measuring the quality of ES, iPS or modified primary cells or cell lines. 

• Samples can be tested under Good Laboratory Practice (GLP) compliance, if required.
• All assays incorporate standards and controls and have been validated according to FDA guidelines for bioanalytical method validation.
• Assay calibration and standardization allows for proficiency testing and for results to be compared between samples over time.
• The ATP bioluminescence technology used in HALO^® QC is a signal detection readout that provides the highest sensitivity available to detect the rarest of stem cell populations.
• HALO^® QC studies are usually completed in just 5 days.
• After completion of the assessment, a report will be sent (usually by email) within 2-3 business days.

Please note that stem cell "quality" should not be mistaken as potency or strength. 

Assessing Umbilical Cord Blood Bankability 

• Only cord blood units demonstrating the highest stem cell "quality" should be permanently stored for clinical use. A rapid, single stem cell assay was developed for this purpose that has the capability of measuring large numbers of cord blood samples with high throughput capability. This assay is called STEMpredict™.
• STEMpredict™ is a unique, 3-day in vitro assay that determines functional metabolic viability (cellular function) and whether the stem cells in the unit have the ability to proliferate.
• STEMpredict™ is usually performed on a sample of the fresh umbilical cord blood, but can also be used on cryopreserved samples.
• STEMpredict™ is capable of triaging low quality from high quality cord blood or mobilized peripheral blood samples. 
• HemoGenix® has determined acceptance values above which stem cell growth functionality can be predicted.

Assessing the Optimum Time for Mobilized Peripheral Blood Collection

• Stem cells are mobilized in patients by consecutive administrations of granulocyte colony-stimulating factor (G-CSF). A patient may or may not "mobilize" or may "mobilize" to a lower degree than expected.
• STEMpredict™ can help determine whether a patient has "mobilized" and the extent of the mobilization".
• The same 3-day STEMpredict™ assay used to determine cord blood bankability can be used to rapidly determine whether stem cells have been induced as a result of G-CSF administration or other mobilizing agent and whether those stem cells can be stimulated to grow.

Samples can be tested under Good Laboratory Practice (GLP) compliance, if required.

• The assay is standardized and validated.
• Results allow samples to be compared over time.
• Proprietary Suspension Expansion Culture™ (SEC™) and ATP bioluminescence technology provides the highest assay sensitivity available with the most reliable results.
• After completion of the study, a report will be sent (usually by email) within 2-3 business days.
• STEMpredict™ is a "best practice criteria testing" platform.

After hematopoietic stem cell transplantation, early phase reconstitution is determined by the ability of the patient to produce red blood cells, granulocytes (neutrophils) and platelets that can be detected in the circulation. The time at which these entities are found can vary significantly, but usually occurs within 2-3 weeks after transplantation. Ahead of significant numbers of cells appearing in the circulation, primitive progenitor cells can be detected using HALO®-96 PMT to measure:

• Erythropoietic progenitor cells (BFU-E),
• Granulocyte-Macrophage progenitor cells (GM-CFC), and
• Megakaryopoietic progenitor cells (Mk-CFC).

Unlike the colony-forming unit (CFU) assay which takes between 12 - 14 days to perform, HALO^®-96 PMT takes only 5 days and provides predictive information regarding the time to engraftment.

For remission to occur after stem cell transplantation, all cell lineages of both the hematopoietic and lymphopoietic systems must be functioning. The immune system usually reconstitutes later than hematopoiesis. However, the latter must also be shown to be tri-lineage reconstitution. To demonstrate that reconstitution is balanced, HemoGenix® provides three types of "global" reconstitution assays.

Global" Tri-Lineage, 4-Population Hematopoietic Reconstitution

This assay detects:

• Primitive hematopoietic stem cell population (CFC-GEMM)
• Erythropoietic progenitor cells (BFU-E),
• Granulocyte-Macrophage progenitor cells (GM-CFC), and
• Megakaryopoietic progenitor cells (Mk-CFC).

"Global", 5-Population Hematopoietic Reconstitution

This assay detects:

• Primitive lympho-hematopoietic stem cell (HPP-SP)
• Primitive hematopoietic stem cell population (CFC-GEMM)
• Erythropoietic progenitor cells (BFU-E)
• Granulocyte-Macrophage progenitor cells (GM-CFC), and
• Megakaryopoietic progenitor cells (Mk-CFC).

"Global" 7-Population Lympho-Hematopoietic Reconstitution

This assay detects the same 5-populations as above, but also includes the T- and B-lymphocyte lineages.

• Primitive lympho-hematopoietic stem cell (HPP-SP)
• Primitive hematopoietic stem cell population (CFC-GEMM)
• Erythropoietic progenitor cells (BFU-E)
• Granulocyte-Macrophage progenitor cells (GM-CFC)
• Megakaryopoietic progenitor cells (Mk-CFC).
• T-Lymphopoietic progenitor cells (T-CFC)
• B-Lymphopoietic progenitor cells (B-CFC)

Patient Monitoring During Clinical Trials

To determine the response of patients to drugs and the occurence of potential toxicity to specific stem and progenitor cell populations during clinical trials, the same "global" HALO^®-96 PMT assays can be employed using a small volume of peripheral blood or bone marrow biopsy, if performed.

Advantages of HALO^®-96 PMT to Determine Time to Engraftment for

"Global" Reconstitution or Patient Monitoring during Clinical Trials

• Like all other HALO^® assays, HALO®-96 PMT is always calibrated and standardized prior to sample measurement. This means that the cell response can be compared directly from one sample to another over time.
• HALO^®-96 PMT measures the ability of individual cell populations to proliferate providing the most rapid and predictive information available.
• Thanks to SEC™ and Bioluminomics™ technology, all assays provide validated, reliable and accurate results, using advanced cell culture and the most sensitive assay readout available.
• Rapid 5 day assay turnaround time that can be extended to 7 days for increased sensitivity.
• For patient studies and clinical trial monitoring, in particular, studies can be performed GLP or non-GLP.

Preferred Cell Systems™has the largest, most accurate and reliable assay kits to test hematopoietic cellular therapy and regenerative medicine products in-house. Please click on the links below to take you to the assay kit page on the Preferred Cell Systems website.

STEMpredict™

HALO^® QC

HALO^® SC-IPS

HALO^® TE

HALO^® PMT

MSCGlo™ QC

MSCGlo™Potency