JavaScript 似乎在您的浏览器中被禁用。 为了您在我们网站能获得最好的体验，建议在浏览器中打开Javascript。

若您需要咨询产品或有任何技术问题，请通过官方电话 400 885 9050 或邮箱 info.cn@stemcell.com 与我们联系

MethoCult™SF M3436

含重组细胞因子（包括EPO）的小鼠红细胞祖细胞无血清甲基纤维素培养基

率先评论此产品

产品号 #（选择产品）

产品号 #03436_C

含重组细胞因子（包括EPO）的小鼠红细胞祖细胞无血清甲基纤维素培养基

跳到结尾的图片库

跳转到图像库的开头

总览

实验数据

产品说明书及文档

应用领域

总览

MethoCult™SF M3436针对小鼠骨髓、胎肝、脾、外周血和纯化细胞的集落形成单位（CFU）检测中原始红细胞祖细胞（BFU-E）的生长和计数进行了优化。MethoCult™SF M3436不支持粒细胞-巨噬细胞祖细胞（CFU-GM、CFU-G和CFU-M）或多潜能粒细胞、红细胞、巨噬细胞、巨核细胞祖细胞（CFU-GEMM）的生长。该制剂不含血清，与STEMvision™软件兼容，用于小鼠骨髓CFU检测的自动菌落计数。MethoCult™SF M3436也可用于大鼠骨髓细胞CFU检测中BFU-E的生长和计数。

浏览我们的常见问题（FAQs）进行CFU化验。

Contains
• Methylcellulose in Iscove's MDM
• Bovine serum albumin
• Recombinant human insulin
• Human transferrin (iron-saturated)
• 2-Mercaptoethanol
• Cytokines (including recombinant human erythropoietin [EPO])
• Supplements

Subtype
Semi-Solid Media, Specialized Media

Cell Type
Hematopoietic Stem and Progenitor Cells

Species
Mouse, Rat

Application
Cell Culture, Colony Assay, Functional Assay

Brand
MethoCult

Area of Interest
Drug Discovery and Toxicity Testing, Stem Cell Biology

Formulation Category
Serum-Free

查看更多显示更少

实验数据

Figure 1. Procedure Summary for Hematopoietic CFU Assays

Figure 2. Examples of Colonies Derived From Mouse Hematopoietic Progenitors

BFU-E-derived colonies from mouse fetal liver (A) and spleen (B) samples cultured in MethoCult™ SF M3436

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

MethoCult™ SF M3436

Catalog #

03436

Lot #

All

Language

English

Document Type

Technical Manual

Product Name

MethoCult™ SF M3436

Catalog #

03436

Lot #

All

Language

English

Document Type

Safety Data Sheet

Product Name

MethoCult™ SF M3436

Catalog #

03436

Lot #

All

Language

English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area

Workflow Stages

Workflow Stages for Hematopoietic Stem and Progenitor Cell Research

Cell Sourcing and Isolation

Expansion and Differentiation

Analysis

Why use semi-solid media?

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.

Why use methylcellulose-based media?

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.

Is it necessary to add antibiotics to the media?

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.

Is there anything I can do if my cultures appear contaminated?

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.

Why can't I use a pipette to dispense methylcellulose-based media?

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™.

Can I 'pluck' the colonies for individual analysis?

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.

Why are low adherence dishes so important?

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

Can MethoCult™ products be used for lymphoid progenitor CFU assays?

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).

Is it possible to set up CFU assays in a 24-well plate?

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.

Can I stain colonies in MethoCult™ medium?

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.

Are there differences in colony morphology with serum-free media?

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.

Can MethoCult™ be made with alternate base media?

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

Is there a MethoCult™ formulation suitable for HPP-CFC (high proliferative potential colony forming cell)?

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.

Genomic subtyping and therapeutic targeting of acute erythroleukemia. I. Iacobucci et al. Nature genetics 2019

Abstract

Acute erythroid leukemia (AEL) is a high-risk leukemia of poorly understood genetic basis, with controversy regarding diagnosis in the spectrum of myelodysplasia and myeloid leukemia. We compared genomic features of 159 childhood and adult AEL cases with non-AEL myeloid disorders and defined five age-related subgroups with distinct transcriptional profiles: adult, TP53 mutated; NPM1 mutated; KMT2A mutated/rearranged; adult, DDX41 mutated; and pediatric, NUP98 rearranged. Genomic features influenced outcome, with NPM1 mutations and HOXB9 overexpression being associated with a favorable prognosis and TP53, FLT3 or RB1 alterations associated with poor survival. Targetable signaling mutations were present in 45{\%} of cases and included recurrent mutations of ALK and NTRK1, the latter of which drives erythroid leukemogenesis sensitive to TRK inhibition. This genomic landscape of AEL provides the framework for accurate diagnosis and risk stratification of this disease, and the rationale for testing targeted therapies in this high-risk leukemia.

View publication

Inhibition of the TGF-beta receptor I kinase promotes hematopoiesis in MDS. Zhou L et al. Blood 2008 OCT

Abstract

MDS is characterized by ineffective hematopoiesis that leads to peripheral cytopenias. Development of effective treatments has been impeded by limited insight into pathogenic pathways governing dysplastic growth of hematopoietic progenitors. We demonstrate that smad2, a downstream mediator of transforming growth factor-beta (TGF-beta) receptor I kinase (TBRI) activation, is constitutively activated in MDS bone marrow (BM) precursors and is overexpressed in gene expression profiles of MDS CD34(+) cells, providing direct evidence of overactivation of TGF-beta pathway in this disease. Suppression of the TGF-beta signaling by lentiviral shRNA-mediated down-regulation of TBRI leads to in vitro enhancement of hematopoiesis in MDS progenitors. Pharmacologic inhibition of TBRI (alk5) kinase by a small molecule inhibitor, SD-208, inhibits smad2 activation in hematopoietic progenitors, suppresses TGF-beta-mediated gene activation in BM stromal cells, and reverses TGF-beta-mediated cell-cycle arrest in BM CD34(+) cells. Furthermore, SD-208 treatment alleviates anemia and stimulates hematopoiesis in vivo in a novel murine model of bone marrow failure generated by constitutive hepatic expression of TGF-beta1. Moreover, in vitro pharmacologic inhibition of TBRI kinase leads to enhancement of hematopoiesis in varied morphologic MDS subtypes. These data directly implicate TGF-beta signaling in the pathobiology of ineffective hematopoiesis and identify TBRI as a potential therapeutic target in low-risk MDS.

View publication

View All Publications

MethoCult™SF M3436

产品号 #（选择产品）

产品号 #03436_C

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

Request Pricing

更多信息

Scientists Helping Scientists™

种属	Mouse, Rat
Contains	• Methylcellulose in Iscove's MDM • Bovine serum albumin • Recombinant human insulin • Human transferrin (iron-saturated) • 2-Mercaptoethanol • Cytokines (including recombinant human erythropoietin [EPO]) • Supplements
配方类别	Serum-Free

MethoCult™SF M3436

产品号 #（选择产品）

产品号 #03436_C

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

技术资料 (8)

常见问题

Why use semi-solid media?

Why use methylcellulose-based media?

Is it necessary to add antibiotics to the media?

Is there anything I can do if my cultures appear contaminated?

Why can't I use a pipette to dispense methylcellulose-based media?

Can I 'pluck' the colonies for individual analysis?

Why are low adherence dishes so important?

Can MethoCult™ products be used for lymphoid progenitor CFU assays?

Is it possible to set up CFU assays in a 24-well plate?

Can I stain colonies in MethoCult™ medium?

Are there differences in colony morphology with serum-free media?

Can MethoCult™ be made with alternate base media?

Is there a MethoCult™ formulation suitable for HPP-CFC (high proliferative potential colony forming cell)?

文献 (2)

Abstract

Abstract

Request Pricing

更多信息

欢迎您关注STEMCELL Technologies的微信公众平台