HemoGenix Inc

Colorado Springs, CO, United States

HemoGenix Inc

Colorado Springs, CO, United States

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Rich I.N.,HemoGenix Inc
Stem Cells Translational Medicine | Year: 2015

This article critically reviews current methods to test and characterize umbilical cord blood (UCB) for hematopoietic stem cell transplantation. These tests include total nucleated cell (TNC) count, viability, viable CD34-positive content, and the colony-forming unit assay. It is assumed that the data obtained are sufficient to perform aUCB stem cell transplantwithout actually determining the quality and potency of the stem cells responsible for engraftment. This assumption has led not only to a high graft failure rate attributed to low or lack of potency, but also to noncompliance with present statutes that require UCB stem cells to be of high quality and, indeed, potency for a transplant to be successful. New evidence now calls into question the quality of the data, based on the UCB processed TNC fraction because using this impure fraction masks and significantly underestimates the functionality of the stem cells in both the segment and the unit. It is proposed that UCB units should be processed to themononuclear cell fraction and that new cost-effective technology thatmeasures the quality and potency of UCB stem cells be implemented to achieve better practices in UCB testing. These changeswould provide the transplant physician with the assurance that the stem cellswill perform as intended and would reduce risk and increase safety and efficacy for the patient. © AlphaMed Press.


Patterson J.,University of Colorado at Colorado Springs | Moore C.H.,University of Colorado at Boulder | Palser E.,University of Colorado at Colorado Springs | Hearn J.C.,HemoGenix Inc | And 3 more authors.
Journal of Translational Medicine | Year: 2015

Rare hematopoietic stem cell populations are responsible for the transplantation engraftment process. Umbilical cord blood (UCB) is usually processed to the total nucleated cell (TNC), but not to the mononuclear cell (MNC) fraction. TNC counts are used to determine UCB unit storage, release for transplantation and correlation with time to engraftment. However, the TNC fraction contains varying concentrations of red blood cells, granulocytes, platelets and other cells that dilute and mask the stem cells from being detected. This does not allow the quality and potency of the stem cells to be reliably measured. Methods: 63 UCB segments and 10 UCB units plus segments were analyzed for the response of both primitive lympho-hematopoietic and primitive hematopoietic stem cells in both the TNC and MNC fractions. The samples were analyzed using a highly sensitive, standardized and validated adenosine triphosphate (ATP) bioluminescence stem cell proliferation assay verified against the colony-forming unit (CFU) assay. Dye exclusion and metabolic viability were also determined. Results: Regardless of whether the cells were derived from a segment or unit, the TNC fraction always produced a significantly lower and more variable stem cell response than that derived from the MNC fraction. Routine dye exclusion cell viability did not correspond with metabolic viability and stem cell response. Paired UCB segments produced highly variable results, and the UCB segment did not produce similar results to the unit. Discussion: The TNC fraction underestimates the ability and capacity of the stem cells in both the UCB segment and unit and therefore provides an erroneous interpretation of the of the results. Dye exclusion viability can result in false positive values, when in fact the stem cells may be dead or incapable of proliferation. The difference in response between the segment and unit calls into question the ability to use the segment as a representative sample of the UCB unit. It is apparent that present UCB processing and testing methods are inadequate to properly determine the quality and potency of the unit for release and use in a patient. © Patterson et al.; licensee BioMed Central.


PubMed | HemoGenix Inc
Type: | Journal: Methods in molecular biology (Clifton, N.J.) | Year: 2014

All stem cells exhibit the capacity and ability to proliferate. This is a fundamental property of stem cells, since it is required not only for self-renewal, but also for expansion and the ability for stem cells to engraft in a patient. The capacity or potential for proliferation by stem cells defines their degree of primitives or stemness. This, in turn, is directly related to stem cell self-renewal. Using a highly sensitive, accurate, and reliable ATP bioluminescence signal detection system that can be multiplexed with other assay readouts, it has been possible to determine three stem cell parameters using a single assay. Using primitive hematopoietic bone marrow stem cells as an example, an in vitro protocol is described that incorporates initially culturing primitive stem cells to induce them into cell cycle, followed by a secondary re-plating step that demonstrates both self-renewal capability and expansion potential.


PubMed | HemoGenix Inc
Type: | Journal: Methods in molecular biology (Clifton, N.J.) | Year: 2014

The potency of a drug is one of the most important parameters of a therapeutic. Besides providing the basis for manufacturing consistency and product stability, the potency can predict product failure or toxicity due to incorrect potency, provide release criteria, and the dose that will ensure that it can be used as intended. Recently, cellular therapeutics, in particular, stem cell therapy products, have being designated as drugs by regulatory agencies if they produce a systemic effect in the patient. Regulatory agencies are becoming increasingly stringent with respect to the manufacture, production, and testing of these products prior to being used in a patient. A clear understanding of what potency is and how it can be measured should help erase the misunderstandings and misconceptions that have accrued within the cellular therapy field. This protocol describes how the potency of hematopoietic stem cell therapy products is determined. The same principles apply to any proliferating stem cell therapeutic product.

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