MesenCult™ Expansion Kit (Mouse)

For the culture of mouse MSCs and MEFs

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MesenCult™ Expansion Kit (Mouse)

For the culture of mouse MSCs and MEFs

1 Kit
Catalog #05513
227 USD

Required Products

Overview

MesenCult™ Expansion Kit (Mouse) is standardized for the culture of mouse mesenchymal stromal cells (MSCs; also known as mesenchymal stem cells) and mouse embryonic fibroblasts (MEFs). The kit includes MesenCult™ Basal Medium (Mouse), MesenCult™ 10X Supplement (Mouse), and MesenPure™. MesenCult™ Expansion Medium has been optimized for the derivation and expansion of mouse MSCs and MEFs in vitro as well as for the detection of colony-forming unit–fibroblasts (CFU­F). This kit was optimized using cells from the mouse strain C57BL/6.
To facilitate the enrichment of MSCs and MEFs during cell culture without serial passaging and frequent medium changes, simply add MesenPure™ to complete MesenCult™ Expansion Medium just prior to use. Although not required, the addition of MesenPure™ is strongly recommended, as the resulting MSC and MEF cultures are more homogeneous and exhibit more robust proliferation, differentiation, and colony formation when compared to complete MesenCult™ Expansion Medium alone.
NOTE: MesenCult™ Expansion Medium must be supplemented with L-Glutamine (Catalog #07100).
Advantages:
• Fast expansion of mouse MSCs with robust enrichment as early as passage 0.
• Optimized for use with mouse bone marrow-, compact bone- and adipose-derived MSCs and MEFs.
• Obtain homogeneous mouse MSC cultures while maintaining tri-lineage differentiation potential.
• Rigorous raw material screening and quality control minimize lot-to-lot variability and increase reproducibility between experiments.
Components:
  • MesenCult™ Basal Medium (Mouse), 450 mL
  • MesenCult™ 10X Supplement (Mouse), 50 mL
  • MesenPure™, 0.5 mL
Subtype:
Specialized Media
Cell Type:
Mesenchymal Stem and Progenitor Cells; Mouse Embryonic Fibroblasts
Species:
Mouse
Application:
Cell Culture; Colony Assay; Expansion
Brand:
MesenCult
Area of Interest:
Stem Cell Biology

Scientific Resources

Educational Materials

(6)

Product Applications

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.

Data and Publications

Data

Procedure Summary for Hematopoietic CFU Assays

Figure 1. CFU-F Assay Comparing Mouse Bone Marrow (BM) MSCs Derived and Cultured in MesenCult™ Expansion Medium With and Without MesenPure™, and Other Commercially Available Media

Numerous CFU-F colonies were observed in cultures maintained in (A) MesenCult™ Expansion Medium (Control) and in (B) same medium containing MesenPure™. Few to no colony formation were observed when cultures were maintained in (C) Commercial Medium 1 or (D) Commercial Medium 2. Seeding density: 5x10^4 cells/cm^2.

Procedure Summary for Hematopoietic CFU Assays

Figure 2. Long-Term Expansion of Mouse BM-Derived MSCs is Observed When Cells are Cultured in MesenCult™ Expansion Medium

Mouse BM MSCs, derived and cultured in MesenCult™ Expansion Medium (Control), show superior long-term expansion rate compared to Commercial Medium 1 and 2. The addition of MesenPure™ enriches for MSCs as early as passage 0 and further improves the expansion rate beyond passage 8. The doubling time of mouse MSCs cultured with or without MesenPure™ are 2.29 and 3.01, respectively. BM MSCs culture-expanded using the MesenCult™ Expansion Kit, with or without MesenPure™, were done under hypoxic conditions. BM MSCs culture-expanded in Commercial Medium 1 and 2 were culture-expanded under normoxic conditions as recommended by their protocols. Data shown from one representative experiment (n=3).

Procedure Summary for Hematopoietic CFU Assays

Figure 3. Mouse BM- and Compact Bone (CB)-Derived MSCs Culture-expanded in MesenCult™ Expansion Medium With or Without MesenPure™ Maintain Multi-Lineage Differentiation Potential

Enriched populations of MSCs were observed at earlier passages upon addition of MesenPure™, which showed increased and more dense differentiation than control cultures. (A) Mouse BM MSCs culture-expanded in MesenCult™ Expansion Medium (Control) differentiated into (B) adipocytes; and (C) osteoblasts. (D) Mouse BM-derived MSCs culture-expanded with MesenPure™ differentiated into (E) adipocytes; and (F) osteoblasts . Differentiation of mouse BM MSCs into chondrocytes is in progress. (G) Mouse CB MSCs culture-expanded in MesenCult™ Expansion Medium (Control) differentiated into (H) adipocytes, (I) osteoblasts and (J) chondrocytes. Adipose-derived mesenchymal stem and progenitor cells, and mouse embryonic fibroblasts (MEFs) were derived and culture-expanded using the MesenCult™ Expansion Kit. These cells were also differentiated towards the adipogenic and osteogenic lineages (data not shown). Adipocytes were stained with Oil Red O staining. Osteoblasts were stained with Alkaline phosphatase and silver nitrate (von Kossa). Chondrocytes were stained with Alcian Blue and Nuclear Fast Red. Images were taken at passage 2.

Procedure Summary for Hematopoietic CFU Assays

Figure 4. Flow Cytometric Analysis of Culture-Expanded Mouse BM-Derived MSCs Using the MesenCult™ Expansion Kit

Mouse BM MSCs were culture-expanded in MesenCult™ Expansion Medium (Control) or with MesenPure™. MSCs from passage 2 were stained for the mesenchymal surface markers, CD106 and Sca1, and the hematopoietic marker, CD45. Stained cells were then analyzed by flow cytometry. MSCs culture-expanded in Control medium show distinct populations of CD45+ hematopoietic cells and CD45- (CD106+ and Sca1+) MSCs. Upon addition of MesenPure™ to the Control Medium, an enriched and homogenous population of CD45- (CD106+ and Sca1+) MSCs are obtained.

Publications

(4)
Stem cells translational medicine 2019 jun

Mesenchymal Stromal Cells Modulate Corneal Alloimmunity via Secretion of Hepatocyte Growth Factor.

S. K. Mittal et al.

Abstract

Mesenchymal stromal cells (MSCs) are multipotent stem cells that participate in tissue repair and posses considerable immunomodulatory potential. MSCs have been shown to promote allograft survival, yet the mechanisms behind this phenomenon have not been fully defined. Here, we investigate the capacity of MSCs to suppress the allogeneic immune response by secreting the pleiotropic molecule hepatocyte growth factor (HGF). Using an in vivo mouse model of corneal transplantation, we report that MSCs promote graft survival in an HGF-dependent manner. Moreover, our data indicate that topically administered recombinant HGF (1) suppresses antigen-presenting cell maturation in draining lymphoid tissue, (2) limits T-helper type-1 cell generation, (3) decreases inflammatory cell infiltration into grafted tissue, and (4) is itself sufficient to promote transplant survival. These findings have potential translational implications for the development of HGF-based therapeutics. Stem Cells Translational Medicine 2019.
Oncotarget 2019 jul

CD90low MSCs modulate intratumoral immunity to confer antitumor activity in a mouse model of ovarian cancer.

Y. Zeng et al.

Abstract

Both anti-tumoral and pro-tumoral effects of mesenchymal stem cells (MSCs) in preclinical treatment of ovarian cancer have been controversially demonstrated. In this study, we profiled the phenotypes of mouse compact bone-derived MSCs (CB-MSCs) and bone marrow-derived MSCs (BM-MSCs) and found that CB-MSCs expressed lower CD90 compared to BM-MSCs. We examined gene expression of immune regulating cytokines of CB-MSCs in 2D and 3D culture and under stimulation with TLR4 agonist LPS or immune activator VIC-008. Our data showed that when CB-MSCs were cultured in simulated in vivo 3D condition, CD90 expression was further decreased. Moreover, gene expressions of immune activating cytokines IL-12, IL-21, IFNgamma and a pro-inflammatory cytokine CXCL10 in CB-MSCs were increased in 3D culture whereas gene expression of anti-inflammatory cytokines IL-10 and CCL5 were downregulated. Stimulation of CB-MSCs by LPS or VIC-008 presented similar profile of the cytokine gene expressions to that in 3D culture which might benefit the anti-tumor efficacy of CD90low MSCs. The anti-tumor effects of CD90low CB-MSCs alone or in combination with VIC-008 were evaluated in a syngeneic orthotopic mouse model of ovarian cancer. Treatment that combines CB-MSCs and VIC-008 significantly decreased tumor growth and prolonged mouse survival. This was associated with the increase of activated anti-tumoral CD4+ and CD8+ T cells and the decrease of Treg cells in the tumor microenvironment. Taken together, our study demonstrates the synergistic anti-tumoral efficacy by application of CB-MSCs combined with immune activator VIC-008 and provides new insight into CD90low MSCs as a new anti-tumor arsenal.
Journal of the American Heart Association 2019 jul

Carnosine Supplementation Mitigates the Deleterious Effects of Particulate Matter Exposure in Mice.

W. Abplanalp et al.

Abstract

Background Exposure to fine airborne particulate matter ( PM 2.5) induces quantitative and qualitative defects in bone marrow-derived endothelial progenitor cells of mice, and similar outcomes in humans may contribute to vascular dysfunction and the cardiovascular morbidity and mortality associated with PM 2.5 exposure. Nevertheless, mechanisms underlying the pervasive effects of PM 2.5 are unclear and effective interventional strategies to mitigate against PM 2.5 toxicity are lacking. Furthermore, whether PM 2.5 exposure affects other types of bone marrow stem cells leading to additional hematological or immunological dysfunction is not clear. Methods and Results Mice given normal drinking water or that supplemented with carnosine, a naturally occurring, nucleophilic di-peptide that binds reactive aldehydes, were exposed to filtered air or concentrated ambient particles. Mice drinking normal water and exposed to concentrated ambient particles demonstrated a depletion of bone marrow hematopoietic stem cells but no change in mesenchymal stem cells. However, HSC depletion was significantly attenuated when the mice were placed on drinking water containing carnosine. Carnosine supplementation also increased the levels of carnosine-propanal conjugates in the urine of CAPs-exposed mice and prevented the concentrated ambient particles-induced dysfunction of endothelial progenitor cells as assessed by in vitro and in vivo assays. Conclusions These results suggest that exposure to PM 2.5 has pervasive effects on different bone marrow stem cell populations and that PM 2.5-induced hematopoietic stem cells depletion, endothelial progenitor cell dysfunction, and defects in vascular repair can be mitigated by excess carnosine. Carnosine supplementation may be a viable approach for preventing PM 2.5-induced immune dysfunction and cardiovascular injury in humans.
Acta neuropathologica 2019 jul

Detrimental and protective action of microglial extracellular vesicles on myelin lesions: astrocyte involvement in remyelination failure.

M. Lombardi et al.

Abstract

Microglia are highly plastic immune cells which exist in a continuum of activation states. By shaping the function of oligodendrocyte precursor cells (OPCs), the brain cells which differentiate to myelin-forming cells, microglia participate in both myelin injury and remyelination during multiple sclerosis. However, the mode(s) of action of microglia in supporting or inhibiting myelin repair is still largely unclear. Here, we analysed the effects of extracellular vesicles (EVs) produced in vitro by either pro-inflammatory or pro-regenerative microglia on OPCs at demyelinated lesions caused by lysolecithin injection in the mouse corpus callosum. Immunolabelling for myelin proteins and electron microscopy showed that EVs released by pro-inflammatory microglia blocked remyelination, whereas EVs produced by microglia co-cultured with immunosuppressive mesenchymal stem cells promoted OPC recruitment and myelin repair. The molecular mechanisms responsible for the harmful and beneficial EV actions were dissected in primary OPC cultures. By exposing OPCs, cultured either alone or with astrocytes, to inflammatory EVs, we observed a blockade of OPC maturation only in the presence of astrocytes, implicating these cells in remyelination failure. Biochemical fractionation revealed that astrocytes may be converted into harmful cells by the inflammatory EV cargo, as indicated by immunohistochemical and qPCR analyses, whereas surface lipid components of EVs promote OPC migration and/or differentiation, linking EV lipids to myelin repair. Although the mechanisms through which the lipid species enhance OPC maturation still remain to be fully defined, we provide the first demonstration that vesicular sphingosine 1 phosphate stimulates OPC migration, the first fundamental step in myelin repair. From this study, microglial EVs emerge as multimodal and multitarget signalling mediators able to influence both OPCs and astrocytes around myelin lesions, which may be exploited to develop novel approaches for myelin repair not only in multiple sclerosis, but also in neurological and neuropsychiatric diseases characterized by demyelination.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.