| PRODUCT | QUANTITY | CATALOG # |
|---|
| MethoCult® Express | 100 mL | 04437 | | 24 x 3 mL | 04447 |
A new and faster assay for CFC enumeration in cord blood To measure the total CFC content of a cord blood unit, it is not necessary to distinguish colony types, and therefore neither is it necessary to wait until colonies have fully matured. MethoCult® Express (Catalog #04437/04447): - is formulated for accelerated progenitor proliferation and colony formation, resulting in larger colonies compared to standard media.
- allows cultures to be counted as early as after 7 days, instead of the 14 - 16 days of conventional CFC assays.
- produces day 7 total CFC numbers in Methocult® Express that show excellent correlation with day 14 total CFC numbers in MethoCult® Express and other MethoCult® media (see images below).
Cultures in MethoCult® Express can be continued for another 7 - 9 days (for a total of 14 - 16 days) after which burst-forming unit-erythroid (BFU-E), CFU-GM and colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM) can be counted separately, as in conventional CFC assays.
Importance of CFC assays Colony-forming cell (CFC) or colony-forming unit (CFU) assays are the most commonly used assays to measure the frequency and proliferative ability of hematopoietic cells in cell preparations used for clinical transplantation. Colony-forming unit-granulocyte, macrophage (CFU-GM) or total CFC numbers in hematopoietic grafts are good predictors of neutrophil and platelet engraftment and survival after transplantation.1-6 In unrelated CB transplantation, total CFC numbers after thawing of CB units correlated more strongly with recovery and survival than other parameters, such as CFC numbers before cryopreservation, TNC counts and CD34+ cell numbers.6 CFC content of attached segments has been shown to correlate well with the CFC content of the whole CB unit.7 Therefore, CFC assays on attached segments can, in principle, be used to measure the CFC content of CB units prior thawing of the whole bag, assuming that proper procedures for freezing and thawing, method validation and quality control are in place. Assessing the potency and quality of hematopoietic cells is particularly important in cord blood (CB) transplantation because: - cell numbers available for transplantation are limited;
- there is large variability in stem cell and progenitor content between different CB units and
- ex vivo manipulations such as red blood cell removal, volume reduction, cryopreservation and thawing of CB units can have large and variable effects on cell yield and viability of the cells.
Advantages of the CFC assay over other assays Other assay formats, which measure the overall metabolic or proliferative activity of a cell preparation, have been proposed as alternatives to CFC assays to measure the quality of candidate CB products. Unlike the CFC Assay, - these other assay formats do not provide information about progenitor frequencies.
- the accuracy and specificity of these assays may be affected by uncontrollable parameters, such as different types and numbers of mature blood cells, nonhematopoietic cells, cellular breakdown products and other compounds in the cell product.
Hematopoietic Colonies Derived from Human Cord Blood Progenitors in
MethoCult® Express Medium After 7 days and 14 days Note that the same fields of view are presented for a side-by-side comparison at 7 and 14 days, showing that colonies after 7 days correlate with colonies after 14 days.
References 1. Migliaccio AR et al.: Cell dose and speed of engraftment in placental/umbilical cord blood transplantation: graft progenitor cells content is a better predictor than nucleated cell quantity. Blood 96:2717-2722, 2000
2. Hogge DE et al.: Quantitation of primitive and lineage-committed progenitors in mobilized peripheral blood for prediction of platelet recovery post autologous transplant. Bone Marrow Transplant 25:589-598, 2000
3. Iori AP et al.: Pre-transplant prognostic factors for patients with high-risk leukemia undergoing an unrelated cord blood tranplantation. Bone Marrow Transplant 33:1097-1105, 2004
4. Yang H et al.: Association of post-thaw viable CD34+ cells and CFU-GM with time to hematopoietic engraftment. Bone Marrow Transplant 35:881-881, 2005
5. Yoo KH et al.: The impact of post-thaw colony-forming units-granulocyte/macrophage on engraftment following unrelated cord blood transplantation in pediatric recipients. Bone Marrow Transplant 39:515-521, 2007
6. Prasad VK et al.: Unrelated donor umbilical cord blood transplantation for inherited metabolic disorders in 159 pediatric patients from a single center: influence of cellular composition of the graft on transplantation outcomes. Blood 112:2979-2989, 2008
7. Rodríguez L, García J, Querol S: Predictive utility of the attached segment in the quality control of a cord blood graft. Biology of Blood and Marrow Transplantation 11:247-251, 2005
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