MethoCult™ H4434 Classic Methylcellulose Medium (Human) | STEMCELL Technologies (2023)

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Product Name

Catalog #

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Document Type

Product Information Sheet

Product Name

MethoCult™ H4434 Classic

Catalog #

04434, 04444

Lot #




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Technical Manual

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MethoCult™ H4434 Classic

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Safety Data Sheet

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04434, 04444

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This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area

Workflow Stages

Workflow Stages for Hematopoietic Stem and Progenitor Cell Research

Cell Sourcing and Isolation

Expansion and Differentiation


Workflow Stages for Cord Blood Banking

Cell Processing

Potency Testing

Workflow Stages for Myeloid Cell Research

Cell Sourcing

Cell Isolation

Cell Characterization

Resources and Publications

Educational Materials (5)


Hematopoietic Stem and Progenitor Cells - Products for Your Research


MethoCult™ Media for Performing Hematopoietic Colony-Forming Unit (CFU) Assays


Identification of Colonies Derived from Human Hematopoietic Progenitors



Applications of the Hematopoietic CFU Assay in Toxicity Testing

Mini Review

Hematopoietic Stem and Progenitor Cells (HSPCs): Isolation, Culture, and Assays

Frequently Asked Questions

Semi-solid media (methylcellulose-based MethoCult™ and collagen-based MegaCult™-C) allow the clonal progeny of a single progenitor cell to remain spatially isolated from other colonies within a culture, so they may be separately identified and counted.

Methylcellulose permits better growth of erythroid colonies than other types of semi-solid support systems (eg. agar) while allowing optimal myeloid colony formation. When appropriate cytokines are present, committed progenitor cells of both erythroid and granulocyte/macrophage lineages (CFU-GM, CFU-G, CFU-M) as well as multi-potential progenitor cells (CFU-GEMM), can be assayed simultaneously in the same culture dish.

No, aseptic technique should be sufficient to maintain sterile cultures. However, antibiotics (eg. Penicillin/Streptomycin) or anti-fungals (eg. Amphotericin B) may be added to the methylcellulose medium if desired.

No, once contamination is visible, it is not possible to rescue the cultures by the addition of antibiotics. Bacteria and yeast inhibit colony formation by depleting nutrients or by releasing toxic substances.

Methylcellulose is a viscous solution that cannot be accurately dispensed using a pipette due to adherence of the medium to the walls of the pipette tip. Blunt-End, 16 Gauge needles (Catalog #28110), in combination with 3 cc Syringes (Catalog #28230) are recommended for accurate dispensing of MethoCult™.

Yes, colonies can be 'plucked' using a pipette with 200 µL sterile pipette tips or using a glass Pasteur pipette with an elongated tip. Individual colonies should be placed in a volume of 25 - 50 µL of medium, and diluted into suitable culture medium for further culture or analysis.

Adherent cells such as fibroblasts can cause inhibition of colony growth and obscure visualization of colonies.

Human lymphoid progenitors (B, NK and T) seem to require stromal support for growth therefore cannot be grown in MethoCult™. Mouse pre-B clonogenic progenitors can be grown in MethoCult™ M3630 (Catalog #03630).

Yes, as long as a plating concentration optimized for the smaller surface area of a well in a 24-well plate (1.9 cm2 as compared to ~9.5 cm2 for a 35 mm dish) is used for these assays. The number of replicate wells required to get an accurate estimation of CFU numbers may also need to be increased.

The cells in individual colonies in MethoCult™ can be stained, eg., for analysis of morphology or phenotype, after they are plucked from the dish and washed free of methylcellulose. Colonies grown in collagen-based MegaCult™-C medium can be used for immunohistochemical or enzymatic staining in situ after dehydration and fixation onto glass slides.

Serum-containing media generally give better overall growth (colonies may appear larger) but there are no large differences in total colony numbers when CFU assays using serum-free media and serum-containing media are compared, provided that identical cytokines are present.

Yes, this can be done as a 'custom' media order. Please contact for more information.

Yes, MethoCult™ H4535 (Catalog #04535) can be used for the HPP-CFC assay as it does not contain EPO. The culture period is usually 28 days. It is not necessary to feed these cultures as growth factors in the medium are present in excess. As HPP-CFCs can be quite large, overplating can be a problem. It is recommended to plate cells at two or more different concentrations.

Publications (63)

BCMA peptide-engineered nanoparticles enhance induction and function of antigen-specific CD8+ cytotoxic T lymphocytes against multiple myeloma: clinical applications. J. Bae et al. Leukemia 2020 jan


The purpose of these studies was to develop and characterize B-cell maturation antigen (BCMA)-specific peptide-encapsulated nanoparticle formulations to efficiently evoke BCMA-specific CD8+ cytotoxic T lymphocytes (CTL) with poly-functional immune activities against multiple myeloma (MM). Heteroclitic BCMA72-80 [YLMFLLRKI] peptide-encapsulated liposome or poly(lactic-co-glycolic acid) (PLGA) nanoparticles displayed uniform size distribution and increased peptide delivery to human dendritic cells, which enhanced induction of BCMA-specific CTL. Distinct from liposome-based nanoparticles, PLGA-based nanoparticles demonstrated a gradual increase in peptide uptake by antigen-presenting cells, and induced BCMA-specific CTL with higher anti-tumor activities (CD107a degranulation, CTL proliferation, and IFN-$\gamma$/IL-2/TNF-$\alpha$ production) against primary CD138+ tumor cells and MM cell lines. The improved functional activities were associated with increased Tetramer+/CD45RO+ memory CTL, CD28 upregulation on Tetramer+ CTL, and longer maintenance of central memory (CCR7+ CD45RO+) CTL, with the highest anti-MM activity and less differentiation into effector memory (CCR7- CD45RO+) CTL. These results provide the framework for therapeutic application of PLGA-based BCMA immunogenic peptide delivery system, rather than free peptide, to enhance the induction of BCMA-specific CTL with poly-functional Th1-specific anti-MM activities. These results demonstrate the potential clinical utility of PLGA nanotechnology-based cancer vaccine to enhance BCMA-targeted immunotherapy against myeloma.

Highly efficient editing of the beta-globin gene in patient-derived hematopoietic stem and progenitor cells to treat sickle cell disease. S. H. Park et al. Nucleic acids research 2019 may


Sickle cell disease (SCD) is a monogenic disorder that affects millions worldwide. Allogeneic hematopoietic stem cell transplantation is the only available cure. Here, we demonstrate the use of CRISPR/Cas9 and a short single-stranded oligonucleotide template to correct the sickle mutation in the beta-globin gene in hematopoietic stem and progenitor cells (HSPCs) from peripheral blood or bone marrow of patients with SCD, with 24.5 ± 7.6{\%} efficiency without selection. Erythrocytes derived from gene-edited cells showed a marked reduction of sickle cells, with the level of normal hemoglobin (HbA) increased to 25.3 ± 13.9{\%}. Gene-corrected SCD HSPCs retained the ability to engraft when transplanted into non-obese diabetic (NOD)-SCID-gamma (NSG) mice with detectable levels of gene correction 16-19 weeks post-transplantation. We show that, by using a high-fidelity SpyCas9 that maintained the same level of on-target gene modification, the off-target effects including chromosomal rearrangements were significantly reduced. Taken together, our results demonstrate efficient gene correction of the sickle mutation in both peripheral blood and bone marrow-derived SCD HSPCs, a significant reduction in sickling of red blood cells, engraftment of gene-edited SCD HSPCs in vivo and the importance of reducing off-target effects; all are essential for moving genome editing based SCD treatment into clinical practice.

Interconversion between Tumorigenic and Differentiated States in Acute Myeloid Leukemia. M. D. McKenzie et al. Cell stem cell 2019 aug


Tumors are composed of phenotypically heterogeneous cancer cells that often resemble various differentiation states of their lineage of origin. Within this hierarchy, it is thought that an immature subpopulation of tumor-propagating cancer stem cells (CSCs) differentiates into non-tumorigenic progeny, providing a rationale for therapeutic strategies that specifically eradicate CSCs or induce their differentiation. The clinical success of these approaches depends on CSC differentiation being unidirectional rather than reversible, yet this question remains unresolved even in prototypically hierarchical malignancies, such as acute myeloid leukemia (AML). Here, we show in murine and human models of AML that, upon perturbation of endogenous expression of the lineage-determining transcription factor PU.1 or withdrawal of established differentiation therapies, some mature leukemia cells can de-differentiate and reacquire clonogenic and leukemogenic properties. Our results reveal plasticity of CSC maturation in AML, highlighting the need to therapeutically eradicate cancer cells across a range of differentiation states.

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What is methylcellulose medium for human cells? ›

MethoCult™ H4434 Classic (MethoCult™ GF H4434) is a complete methylcellulose-based medium for the growth and enumeration of hematopoietic progenitor cells in colony-forming unit (CFU) assays of human bone marrow, mobilized peripheral blood, peripheral blood, and cord blood samples.

What is semi solid methylcellulose based medium? ›

Methylcellulose medium is a type of semi-solid medium that can be used for culturing hematopoietic stem and progenitor cells in colony-forming assays, or culturing hybridomas and Chinese hamster ovarian cells for cell line development.

What does methylcellulose do to your body? ›

This medication is used to treat constipation. It increases the bulk in your stool, an effect that helps to cause movement of the intestines. It also works by increasing the amount of water in the stool, making the stool softer and easier to pass.

What is methylcellulose side effects? ›

Side effects that you should report to your care team as soon as possible:
  • Allergic reactions—skin rash, itching, hives, swelling of the face, lips, tongue, or throat.
  • Choking—chest pain, trouble swallowing or breathing, vomiting.

Is methylcellulose good or bad? ›

Methyl cellulose is considered safe for all animal species. Setting a maximum content in complete diets is not considered necessary. The use of methyl cellulose in animal nutrition is of no concern for consumer safety.

Does methylcellulose cause inflammation? ›

Although the FDA has approved methylcellulose for consumption, it's best to have it in moderation, or even not at all. Your body isn't used to highly processed foods and compounds, so a high intake can easily lead to unwanted inflammation as your body tries to fight off the unrecognized nutrients.

How often can I use methylcellulose? ›

Methylcellulose is usually taken 1 to 3 times per day. Use exactly as directed on the label, or as prescribed by your doctor. Do not use in larger or smaller amounts or for longer than recommended. Overuse of a laxative may cause damage to the nerves, muscles, or tissues in your intestines.

Who should not take Citrucel? ›

Caution is advised if you have diabetes, phenylketonuria (PKU), or any other condition that requires you to limit/avoid these substances in your diet. Ask your doctor or pharmacist about using this product safely. Tell your doctor if you are pregnant before using this medication.

How long does it take for Citrucel to work? ›

It may take 1 to 3 days before this medication starts working. Use this medication regularly to get the most benefit from it. To help you remember, take it at the same time(s) each day. Do not take this medication for more than 7 days unless directed by your doctor.

What are the side effects from Citrucel? ›

In particular, bulk-forming fiber supplements like Citrucel can cause issues such as bloating, nausea, vomiting, and diarrhea in some people ( 1 ). According to the manufacturer, taking Citrucel without enough fluids can also cause the powder to swell up in your throat and pose a choking risk (2).

Does Citrucel work better than Metamucil? ›

Citrucel (methylcellulose) is mainly insoluble fibers that are nonfermentable, so it's less likely to contribute to bloating and gas. Psyllium husk (Metamucil and Konsyl) is rich in both soluble and insoluble fiber. Generally, fiber supplements with mainly insoluble fiber may be a better option for constipation.

Does methylcellulose cause weight loss? ›

Conclusion: The data indicate that HPMC not only reduces body weight, but also normalizes the metabolic abnormalities associated with obesity and suggest that the effects of HPMC on glucose and lipid homeostasis in B6 mice are mediated by improvements in leptin sensitivity resulting from reduced fat absorption.

Is it safe to take methylcellulose every day? ›

There's no evidence that daily use of fiber supplements — such as psyllium (Metamucil, Konsyl, others) or methylcellulose (Citrucel) — is harmful. Fiber has a number of health benefits, including normalizing bowel function and preventing constipation.

What is methylcellulose in cell culture? ›

Methyl cellulose is used to create semi-solid matrices, as a cell culture gel, for culture of colony forming cells such as CFU-G; CFU-GM; cells and other cells used in toxicity assays. Low viscosity, not for plaquing assays or cloning.

What is methylcellulose used for in biology? ›

Methyl cellulose is used as a buffer additive in capillary electrophoresis to control electroosmotic flow for improved separations.

What is the use of methylcellulose in examining live specimens? ›

The use of methylcellulose avoids artifacts of conventional negative stain-TEM by (1) restricting interactions between the nanoparticles, (2) inhibiting binding to the specimen support films and (3) reducing compression after drying.

What medium is used for cell culture? ›

DMEM. Dulbecco's Modified Eagle Medium (DMEM), a widely used cell culture medium, is more nutrient-rich than MEM and is proven for use with many mammalian cell types.

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