MethoCult™ M3434 Methylcellulose-Based Medium (Mouse) | STEMCELL Technologies (2023)

Protocols and Documentation

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

Document Type

Product Name

Catalog #

Lot #


Document Type

Product Information Sheet

Product Name

MethoCult™ GF M3434

Catalog #

03434, 03444

Lot #




Document Type

Technical Manual

Product Name

MethoCult™ GF M3434

Catalog #


Lot #




Product Name

MethoCult™ GF M3434

Catalog #

03434, 03444

Lot #





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


Resources and Publications

Educational Materials (6)


Hematopoietic Stem and Progenitor Cells - Products for Your Research


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


STEMvision™ Automated and Standardized Counting of Mouse Bone Marrow CFU Assays

Technical Bulletin

MethoCult™ Formulation Optimized for Mouse BFU-E Detection


Identification of Colonies Derived from Mouse Hematopoietic Progenitors

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 (167)

Restraining Lysosomal Activity Preserves Hematopoietic Stem Cell Quiescence and Potency. R. Liang et al. Cell stem cell 2020


Quiescence is a fundamental property that maintains hematopoietic stem cell (HSC) potency throughout life. Quiescent HSCs are thought to rely on glycolysis for their energy, but the overall metabolic properties of HSCs remain elusive. Using combined approaches, including single-cell RNA sequencing (RNA-seq), we show that mitochondrial membrane potential (MMP) distinguishes quiescent from cycling-primed HSCs. We found that primed, but not quiescent, HSCs relied readily on glycolysis. Notably, in vivo inhibition of glycolysis enhanced the competitive repopulation ability of primed HSCs. We further show that HSC quiescence is maintained by an abundance of large lysosomes. Repression of lysosomal activation in HSCs led to further enlargement of lysosomes while suppressing glucose uptake. This also induced increased lysosomal sequestration of mitochondria and enhanced the competitive repopulation ability of primed HSCs by over 90-fold in vivo. These findings show that restraining lysosomal activity preserves HSC quiescence and potency and may be therapeutically relevant.

The NAD-Booster Nicotinamide Riboside Potently Stimulates Hematopoiesis through Increased Mitochondrial Clearance. N. Vannini et al. Cell stem cell 2019 mar


It has been recently shown that increased oxidative phosphorylation, as reflected by increased mitochondrial activity, together with impairment of the mitochondrial stress response, can severely compromise hematopoietic stem cell (HSC) regeneration. Here we show that the NAD+-boosting agent nicotinamide riboside (NR) reduces mitochondrial activity within HSCs through increased mitochondrial clearance, leading to increased asymmetric HSC divisions. NR dietary supplementation results in a significantly enlarged pool of progenitors, without concurrent HSC exhaustion, improves survival by 80{\%}, and accelerates blood recovery after murine lethal irradiation and limiting-HSC transplantation. In immune-deficient mice, NR increased the production of human leucocytes from hCD34+ progenitors. Our work demonstrates for the first time a positive effect of NAD+-boosting strategies on the most primitive blood stem cells, establishing a link between HSC mitochondrial stress, mitophagy, and stem-cell fate decision, and unveiling the potential of NR to improve recovery of patients suffering from hematological failure including post chemo- and radiotherapy.

Sequentially inducible mouse models reveal that Npm1 mutation causes malignant transformation of Dnmt3a-mutant clonal hematopoiesis. M. A. Loberg et al. Leukemia 2019 jul


Clonal hematopoiesis (CH) is a common aging-associated condition with increased risk of hematologic malignancy. Knowledge of the mechanisms driving evolution from CH to overt malignancy has been hampered by a lack of in vivo models that orthogonally activate mutant alleles. Here, we develop independently regulatable mutations in DNA methyltransferase 3A (Dnmt3a) and nucleophosmin 1 (Npm1), observed in human CH and AML, respectively. We find Dnmt3a mutation expands hematopoietic stem and multipotent progenitor cells (HSC/MPPs), modeling CH. Induction of mutant Npm1 after development of Dnmt3a-mutant CH causes progression to myeloproliferative disorder (MPD), and more aggressive MPD is observed with longer latency between mutations. MPDs uniformly progress to acute myeloid leukemia (AML) following transplant, accompanied by a decrease in HSC/MPPs and an increase in myeloid-restricted progenitors, the latter of which propagate AML in tertiary recipient mice. At a molecular level, progression of CH to MPD is accompanied by selection for mutations activating Ras/Raf/MAPK signaling. Progression to AML is characterized by additional oncogenic signaling mutations (Ptpn11, Pik3r1, Flt3) and/or mutations in epigenetic regulators (Hdac1, Idh1, Arid1a). Together, our study demonstrates that Npm1 mutation drives evolution of Dnmt3a-mutant CH to AML and rate of disease progression is accelerated with longer latency of CH.

View All Publications


What is MethoCult? ›

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 is a CFU assay? ›

The CFU assay is a hematopoietic functional assay, which is often used to measure the function or potency of hematopoietic progenitors present in stem cell products.

Is methylcellulose toxic? ›

Unlike other naturally derived binders, methylcellulose is allergen-free. It's also gluten-free, non-toxic, flavorless, colorless, and odorless, making it an excellent gelling agent for various foods. Most methylcellulose will pass through the body undigested.

Is methylcellulose safe for daily use? ›

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.

Is methylcellulose all natural? ›

Although methylcellulose comes from the cellulose of plants and vegetables, its development process isn't entirely natural; it's a synthetically made compound with a complex and refined method. Like regular cellulose, methylcellulose isn't digestible because we lack cellulase, the necessary enzyme for digesting it.

Is a higher CFU count better? ›

Oftentimes, the higher the CFUs per serving, the higher the price because the belief is that the higher the CFU count, the better. This is NOT correct! Potency isn't just a number. When choosing a probiotic supplement, there's more to consider than just the number of CFUs per serving.

How many CFUs are recommended? ›

Thus, for individuals in relatively good health, a daily dose of 10-20 billion CFU is likely sufficient for supporting everyday immune and digestive support.

What range of CFU is acceptable? ›

Heterotrophic plate count levels in potable water should be <500 CFU/mL. These levels may increase on occasion, but counts consistently >500 CFU/mL would indicate a general decrease in water quality.

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.

Is methylcellulose a carcinogen? ›

Methylcellulose is the active ingredient in many laxatives, but animal studies indicate that the additive may promote colorectal cancer at levels typically present in processed foods.

Is methylcellulose good or bad? ›

According to the U.S. National Library of Medicine, methylcellulose is approved by both the U.S. Food and Drug Administration and the European Union as safe for human consumption and has no specified limitations regarding use, as there are no observed adverse effects when consumed in moderation.

What is methylcellulose made out of? ›

Methyl cellulose is made from natural cellulose with white to milk-white colour. It is a tasteless powder (Fig. 8.32). It has multiple functions such as film-forming, emulsifiability, adhesive property and thickening property and is widely used in baked goods, fried foods, desserts, candies and soups as food additives.

Is methylcellulose made from corn? ›

Hydroxypropyl methylcellulose is added to dietary supplements as a regulator and thickener to regulate the consistency of the product. Pharma Nord's hydroxypropyl methylcellulose is non-GMO. It is made synthetically and is guaranteed free of, for example, wheat, corn and soy as well as derivatives there of.

Is methylcellulose Vegan? ›

With the unique functionality of methylcellulose, no egg or other animal-based ingredient needs to be added to the finished product. In addition to being vegan, one can also create both an egg-free and allergen-free food with the best bite.

Is methylcellulose a dairy? ›

Methyl Cellulose is dairy free. Methyl Cellulose does not contain milk and should be safe for those with a milk allergy. Fig's dietitian team reviewed this note on methyl cellulose.

Does cellulose cause inflammation? ›

Dietary cellulose induces anti-inflammatory immunity and transcriptional programs via maturation of the intestinal microbiota - PMC.

What is another name for methylcellulose? ›

Methylcellulose is available over-the-counter (OTC) and as a generic. Methylcellulose is available under the following different brand names: Citrucel.

What is the #1 probiotic? ›

The probiotic strain in Culturelle® - Lactobacillus rhamnosus GG (LGG®) – is the #1 most clinically studied strain of probiotic. Considered the premier probiotic in the world, over 1,000 scientific studies and over 30 years of research have demonstrated the safety and efficacy of Lactobacillus rhamnosus GG (LGG®).

What are the signs that probiotics are working? ›

But there are some common indicators that they may be working for you.
  • Less stomach pain. For some people, certain probiotics can help with stomach pain and cramps. ...
  • More frequent poops. ...
  • Less bloating. ...
  • Improved sleep. ...
  • Better mood, memory, and mental clarity. ...
  • Fewer vaginal infections.
Feb 13, 2023

Is 80 billion CFU too much? ›

Probiotic doses are measured by colony-forming units (CFUs), and they range from 1 billion to 100 billion CFUs. Perlmutter considers 100 billion to be the highest daily dosage anybody needs to take — and most people need much less than that.

Is 50 billion probiotics too much? ›

Many healthy adults can safely take upwards up 30-50 billion CFU if they have a reason or desire to. Interestingly, a study published in the journal of Gastroenterology and Hepatology in 2015 showed that between 10 – 20 Billion CFU is all that is really needed.

Who should not take probiotics? ›

Although probiotics are generally safe to use, findings of a review from 2017 suggest that children and adults with severe illnesses or compromised immune systems should avoid using probiotics. Some people with these conditions have experienced bacterial or fungal infections as a result of probiotic use.

Is $100 million probiotic enough? ›

The American Academy of Family Physicians recommends that adults consume between 10 and 20 billion CFUs per day. This is for someone who wants to support balance in their digestive tract. A person experiencing specific gastrointestinal issues may need more as directed by a physician.

How many CFU is considered infection? ›

The diagnosis of catheter-associated urinary tract infection can be made when the urine culture shows 100 or more CFU per mL of urine from a catheterized patient.

Can you take too many CFU? ›

Taking too many probiotics can cause mild discomfort

Taking more than a usual dose — 1 to 10 billion colony forming units (CFUs) — of probiotics doesn't necessarily mean better results and, instead, might provoke some mildly uncomfortable side effects.

How many CFU should a woman have? ›

Women are generally advised to look for a probiotic supplement that contains at least 20-50 billion colony forming units (CFUs) of bacteria. However, in the early stages, a minimum of 1 billion CFUs may be consumed. A wide range of women-specific formulations includes additional ingredients to promote overall health.

What are examples of semi-solid media? ›

Examples of semi-solid media are: Hugh and Leifson's oxidation fermentation medium, Stuart's and Amies media, and Mannitol motility media. Liquid media: These media do not contain any traces of solidifying agents, such as agar or gelatin, and large growth of bacterial colonies can be observed in the media.

What is the difference between semi-solid and solid media? ›

Solid medium contains 1.5 to 2.0 % solidification agent while semi-solid medium contain 0.2 to 0.5 % solidifying agent. When poured into plates, solid medium solidifies and provides a solid surface to grow microorganisms. Semi-solid medium is soft, and it doesn't solidify completely as solid media.

Is methylcellulose in food vegan? ›

With the unique functionality of methylcellulose, no egg or other animal-based ingredient needs to be added to the finished product. In addition to being vegan, one can also create both an egg-free and allergen-free food with the best bite.

How do you make semi-solid agar medium? ›

Suspend 12 g in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Dispense in tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.

When would you use a semisolid medium? ›

Semisolid media are prepared with lower agar concentrations of 0.2 to 0.5%. They have a soft, custard-like consistency and are used to cultivate microaerophilic bacteria or determine bacterial motility by cultivation in stab tubes. Semisolid media can be used to distinguish between typhoid and colon bacilli.

What are the 3 main types of microbiological culture media? ›

Depending upon the addition and quantity of this substance, media are of three types:
  • 3.3.1 Liquid (Broth) Media. Liquid (Broth) Media, such as nutrient broth , tryptic soy broth or glucose broth which are prepared without the use of agar agar. ...
  • 3.3. 2 Semisolid Media. ...
  • 3.3. 3 Solid Media.
Sep 19, 2020

What is the advantage of solid media? ›

The advantage of solid media (tubed or in plates) is that they enable the detection of mixed cultures and contaminants. Egg-based and agar-based media may be used. The main advantage of an egg-based medium is that it best supports the growth of M. tuberculosis and permits niacin testing.

What is the advantage of using liquid media instead of solid media? ›

Solid media is characterized by agar and other solidifying agents, while liquid media contains specific nutrients but has no gelling agents. Some of the advantages of liquid media include; Liquid media offers a uniform culture condition for the growth of bacteria.

What are the disadvantages of liquid media? ›

Answer: Disadvantages of liquid culture- Bacteria that grow in liquid media may not have specific characteristics. Difficult to isolate different types of bacteria from mixed populations. Bacteria grow diffusely in liquids they produce discrete visible growth in solid media.

What is thermal death rate? ›

Thermal death time is how long it takes to kill a specific bacterium at a specific temperature. It was originally developed for food canning and has found applications in cosmetics, producing salmonella-free feeds for animals (e.g. poultry) and pharmaceuticals.

What is methylcellulose in fake meat? ›

The methylcellulose conundrum

Used by high-profile brands like Impossible Foods and Beyond Meat, methylcellulose, or E461, provides attractive gelling and emulsifying properties that boost juiciness and hold plant-based burgers together as they cook.

What is the difference between semi solid and liquid media? ›

The key difference between solid and liquid media is that solid media contain agar while liquid media do not contain agar. That is, agar is a solidification agent in growth media, and solid media contain a solidifying agent while liquid media lack a solidifying agent.

What is the difference between synthetic and non synthetic media? ›

Synthetic media are media that are partially or fully generated by computers. Non-synthetic media can be any other media, i.e., media that is produced by human input. Take a newspaper article, for instance. Written entirely by a human, it clearly falls under non-synthetic media.

What is the purpose of agar in a solid medium? ›

The addition of agar-agar (a complex carbohydrate extracted from seaweed) results in a solid medium. Agar is an ideal solidifying agent for microbiological media because of its melting properties and because it has no nutritive value for the vast majority of bacteria.

Top Articles
Latest Posts
Article information

Author: Rubie Ullrich

Last Updated: 12/27/2022

Views: 5753

Rating: 4.1 / 5 (72 voted)

Reviews: 87% of readers found this page helpful

Author information

Name: Rubie Ullrich

Birthday: 1998-02-02

Address: 743 Stoltenberg Center, Genovevaville, NJ 59925-3119

Phone: +2202978377583

Job: Administration Engineer

Hobby: Surfing, Sailing, Listening to music, Web surfing, Kitesurfing, Geocaching, Backpacking

Introduction: My name is Rubie Ullrich, I am a enthusiastic, perfect, tender, vivacious, talented, famous, delightful person who loves writing and wants to share my knowledge and understanding with you.