MesenCult™ Expansion Kit (Mouse)
For the culture of mouse MSCs and MEFs

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Overview
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).
Data Figures

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.

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).

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.

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.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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.
Resources and Publications
Educational Materials (5)
Publications (16)
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Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.