MethoCult® Express

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Methylcellulose Medium for 7-Day Colony-Forming Cell Assays of Human Hematopoietic Cells

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  • MethoCult® Express 100 mL
  • Label for MethoCult® Express 100 mL
  • Tube label for MethoCult® Express 3 mL
MethoCult® Express 100 mL
MethoCult® Express is recommended for the detection and quantification of human hematopoietic progenitors in cord blood (CB) samples using CFC assays after a minimum culture period of seven days. It is recommended for red blood cell (RBC) depleted CB samples, whole CB samples that have been cryopreserved and thawed, and CB mononuclear cells.
 
MethoCult® Express is optimized for the detection and counting of human hematopoietic progenitor cells after much shorter periods than the 14 - 16 days of standard CFC assays. In MethoCult® Express, colonies containing at least 20 cells can be counted as early as after seven days of culture. At this time, most colonies are immature and have not yet differentiated into morphologically distinguishable colony types. Therefore the colonies counted after seven days of culture give information about the total frequency of hematopoietic progenitor cells present in the sample without distinction between different progenitor types.
If MethoCult® Express cultures are maintained for 14 - 16 days, colonies derived from erythroid progenitors (BFU-E), granulocyte-macrophage progenitors (CFU-GM, CFU-M, CFU-G), and multi-potential granulocyte, erythroid, macrophage and megakaryocyte progenitors (CFU-GEMM) can be counted.
Product Name Description Catalog # Size Price Quantity
MethoCult® Express Pre-aliquoted methylcellulose medium for 7-day colony-forming cell assays of human hematopoietic cells 04447 24 x 3 mL 809.00 USD      
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MethoCult® Express Methylcellulose medium for 7-day colony-forming cell assays of human hematopoietic cells 04437 100 mL 610.00 USD      
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Recommended for:
Detection of human clonogenic hematopoietic progenitor cells from cord blood using CFC assays after a minimum culture period of seven days
Accessory Products:
• Iscove's Modified Dulbecco's Medium (IMDM) with 2% Fetal Bovine Serum (Catalog #07700)
• Blunt-end Needles (Catalog #28110/28120)
• 3 cc Syringe (Catalog #28230/28240)
• 35 mm Culture Dishes (Catalog #27100/27150)
• 60 mm Gridded Scoring Dishes (Catalog #27500)
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:
• Methylcellulose in Iscove's MDM
• Fetal Bovine Serum
• Bovine Serum Albumin
• Cytokines, including erythropoietin (EPO)
• Supplements
Product Type: Specialized cell culture media
Application: Colony assays & quantification
Area of Interest: Cord blood banking, Stem cell biology, Transplantation
Cell Type: Hematopoietic stem & progenitor cells
Medium Type: Methylcellulose-based
Popular Product Line: MethoCult
Species: Human

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • Q. IS IT POSSIBLE TO DISTINGUISH DIFFERENT COLONY TYPES AFTER 7 DAYS OF CULTURE?

    A. No. Although some erythroid colonies can be identified after 7 days of culture on the basis of morphology, attempts to count erythroid and myeloid colonies separately at day 7 tend to underestimate the number of erythroid colonies, and overestimate the number of myeloid colonies detectable after 14 days. Most colonies remain undifferentiated after 7 days of culture and immature erythroid colonies consist of large nonhemoglobinized erythroblasts that are not easily distinguishable from non-erythroid cells.

  • Q. I ONLY WANT TO EVALUATE CFU-GM COLONIES. CAN THAT BE DONE IN A 7-DAY ASSAY IN METHOCULT®EXPRESS, OR DO I NEED A DIFFERENT MEDIUM?

    A. It is not possible to distinguish colony types after 7 days of culture in Methocult® Express (see question above). In principle, a custom medium without EPO would selectively promote the CFU-GM present to form colonies. However, some BFU-E can develop into erythroid colonies after 7 days in the absence of EPO so that a proportion of total colonies detected after 7 days are derived from erythroid progenitors, even in the absence of EPO. This is due to the fact that the survival and first rounds of proliferation of erythroid progenitors, in particular immature BFU-E, is not dependent on EPO. The presence of EPO is only essential for the survival, proliferation and differentiation in later stages of development of hemoglobinized BFU-E colonies.

Background References:

  1. Vinod Prasad 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 (7) 2979-2989 (June 27, 2008)
  2. K H Yoo 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 (9) 515-521 (May 2007)
  3. H Yang et al. Association of post-thaw viable CD34+ cells and CFU-GM with time to hematopoietic engraftment.Bone Marrow Transplant 35 (9) 881-887 (May 2005)
  4. Luciano Rodriguez et al. Predictive utility of the attached segment in the quality control of a cord blood graft.Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 11 (4) 247-251 (April 2005)
  5. A P Iori et al. Pre-transplant prognostic factors for patients with high-risk leukemia undergoing an unrelated cord blood transplantation.Bone Marrow Transplant 33 (11) 1097-1105 (June 2004)
  6. A R Migliaccio et al. Cell dose and speed of engraftment in placental/umbilical cord blood transplantation: graft progenitor cell content is a better predictor than nucleated cell quantity.Blood 96 (8) 2717-2722 (October 15, 2000)
  7. D E Hogge 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 (6) 589-598 (March 2000)
  8. E Conneally et al. Rapid and efficient selection of human hematopoietic cells expressing murine heat-stable antigen as an indicator of retroviral-mediated gene transfer.Blood 87 (2) 456-464 (January 15, 1996)
  9. A M Farese et al. Acceleration of hematopoietic reconstitution with a synthetic cytokine (SC-55494) after radiation-induced bone marrow aplasia.Blood 87 (2) 581-591 (January 15, 1996)

Product Name

Description

Catalog #
HetaSep™ For Isolating Human Nucleated Cells from Peripheral Blood 07806
Ficoll-Paque™ PLUS Density Gradient Medium for the Isolation of Mononuclear Cells 07907
Ficoll-Paque™ PREMIUM Density Gradient Medium for the Isolation of Mononuclear Cells 07908
Ammonium Chloride Solution Reagent for the Lysis of Red Blood Cells 07800

Colonies in MethoCult® Express


Colonies in MethoCult® Express
Colonies derived from cord blood progenitors in MethoCult® Express at (A) day 7 and (B) day 14 with a 2X objective lens; at (C) day 7 and (D) day 14 with a 4X objective lens


Correlation between colony numbers after Day 7 with MethoCult® Express and Day 14 with MethoCult® H4034 Optimum (Catalog #04034/04044) using cryopreserved whole cord blood


Correlation between colony numbers after Day 7 with MethoCult® Express and Day 14 with MethoCult® H4034 Optimum (Catalog #04034/04044) using cryopreserved whole cord blood
Cryopreserved whole cord blood samples (n = 5) were thawed and set up in MethoCult® Express and MethoCult® GF H4034 without further processing. Colonies in MethoCult® Express were counted after 7 days and colonies in MethoCult® GF H4034 were counted after 14 days. Correlation coefficient: 0.973; p = 0.005 (Pearson Product Moment Correlation).


Correlation between colony numbers after Day 7 with MethoCult® Express and Day 14 with MethoCult® H4034 Optimum (Catalog #04034/04044).


Correlation between colony numbers after Day 7 with MethoCult® Express and Day 14 with MethoCult® H4034 Optimum (Catalog #04034/04044).
Samples included cryopreserved whole cord blood (5), cryopreserved cord blood with red blood cells depleted by HetaSep™ (3), fresh cord blood with red blood cells depleted by HetaSep™ (3) and cryopreserved cord blood mononuclear cells isolated by density sedimentation over Ficoll-Paque™ PLUS (3). Samples (n = 14) were set up in MethoCult® Express and MethoCult® GF H4034 and colonies were counted after 7 and 14 days, respectively. Correlation coefficient: 0.997; p <0.0001 (Pearson Product Moment Correlation).


Correlation between colony numbers after Day 7 and Day 14 with MethoCult® Express


Correlation between colony numbers after Day 7 and Day 14 with MethoCult® Express
Samples included cryopreserved whole cord blood samples (5), cryopreserved cord blood with red blood cells depleted by HetaSep™ (3), fresh cord blood with red blood cells depleted by HetaSep™ (3), cryopreserved cord blood mononuclear cells isolated by density sedimentation over Ficoll-Paque™ PLUS (11) and fresh cord blood mononuclear cells isolated by density sedimentation over Ficoll-Paque™ PLUS (2). Samples (n = 24) were set up in MethoCult® Express and colonies were counted after 7 and 14 days. Correlation coefficient: 0.991; p <0.0001 (Pearson Product Moment Correlation).


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