MesenCult™ Proliferation Kit with MesenPure™ (Mouse)

For detection of CFU-F and expansion of mouse mesenchymal stem cells

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MesenCult™ Proliferation Kit with MesenPure™ (Mouse)

For detection of CFU-F and expansion of mouse mesenchymal stem cells

1 Kit
Catalog #05512
263 CAD

Required Products

Overview

MesenCult™ Proliferation Kit with MesenPure™ (Mouse) has been optimized for the enrichment and expansion of mouse bone marrow- and compact bone-derived mesenchymal stem and progenitor cells (MSCs). The addition of MesenPure™ to complete MesenCult™ medium results in cultures that are more homogeneous and exhibit more robust proliferation, differentiation, and colony formation in the CFU-F assay when compared to complete MesenCult™ medium alone. Undesired hematopoietic cells in mouse MSC cultures are depleted without serial passaging and frequent medium changes. Components are pre-screened to minimize lot-to-lot variability.
Advantages:
• Fast enrichment and expansion of mouse MSCs without serial passaging
• Obtain homogeneous mouse MSC cultures with robust self-regeneration and differentiation potential
• Optimized for use with bone marrow- and compact bone-derived mouse MSCs
• Easy to use as MesenPure™ is simply added to complete MesenCult™ medium just prior to use
• Can also be used for enrichment and expansion of mouse embryonic fibroblasts (MEFs)
Components:
  • MesenCult™ Proliferation Kit with MesenPure™ (Mouse, Catalog #05512)
    • MesenCult™ MSC Basal Medium (Mouse), 400 mL (Catalog #05501)
    • MesenCult™ Stem Cell Stimulatory Supplements (Mouse), 100 mL (Catalog #05502)
    • MesenPure™ 1000X, 0.5 mL (Catalog #05500)
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

Technical Resources

Educational Materials

(10)

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

MSC cultures exposed to MesenPure™ appear homogeneous and mostly devoid of hematopoietic cells as early as passage 0 (P0)

Figure 1. MSC cultures exposed to MesenPure™ appear homogeneous and mostly devoid of hematopoietic cells as early as passage 0 (P0)

Primary BM-derived MSCs were cultured in complete MesenCult™ medium without (Control) or with MesenPure™. In control conditions, hematopoietic cells remained in high numbers as late as P2, with many hematopoietic cells still present at P3 (left panels). In MesenPure™-exposed cultures, low numbers of hematopoietic cells were observed as early as P0, with subsequent passages reducing the number further (right panels).

Flow cytometry analysis of P0 MSC culture exposed to MesenPure™ demonstrates significant enrichment of CD45 - /CD29 + /Sca1 + cells

Figure 2. Flow cytometry analysis of P0 MSC culture exposed to MesenPure™ demonstrates significant enrichment of CD45-/CD29+/Sca1+ cells

Primary BM-derived MSCs were cultured for 14 days in complete MesenCult™ medium without (Control) or with MesenPure™ prior to flow cytometry analysis being performed. ~9% of the control culture and ~70% of the MesenPure™-exposed culture was CD45 -. Analysis of the CD45 - population revealed ~75% of the control cells and ~96% of the MesenPure™-exposed cells co-expressed CD29 and Sca1.

Exposure of MSC cultures to MesenPure™ leads to enrichment of CD45- cells and may increase expansion of CD45-/CD29+/Sca1+ cells

Figure 3. Exposure of MSC cultures to MesenPure™ leads to enrichment of CD45- cells and may increase expansion of CD45-/CD29+/Sca1+ cells

Flow cytometry analysis of BM-derived MSC cultures exposed to MesenPure™ shows a marginal increase in the number of CD45- cells, but a 0.25-fold increase in CD45- /CD29+ /Sca1+ cells compared to cultures without MesenPure™. The ratio of CD45- cells and CD45- /CD29+ /Sca1+ cells was applied to the total number of cells to obtain the actual number of cells in culture. Fold-induction from control cultures is reported in parenthesis.

CFU-F assay cultures exposed to MesenPure™ are homogeneous and devoid of hematopoietic cells

Figure 4. CFU-F assay cultures exposed to MesenPure™ are homogeneous and devoid of hematopoietic cells

CFU-F assays were performed with primary BM-derived MSCs. Cells were cultured in complete MesenCult™ medium without (Control) or with MesenPure™ for 14 days, fixed, and stained with toluidine blue. (A) In the control condition, a large number of hematopoietic cells (red arrows) are present and surround the MSCs (yellow arrows). (B) Cultures exposed to MesenPure™ are homogeneous and devoid of hematopoietic cells.

BM-MSCs

Figure 5. BM-MSCs

CB-MSCs

Figure 6. CB-MSCs

In CFU-F assays, exposure of primary cultures of BM- or CB-derived MSCs to MesenPure™ led to an overall minimum 20% increase in the number of MSC colonies observed, independent of the number of cells initially plated.

MSCs exposed to MesenPure™ require fewer cells to demonstrate robust differentiation

Figure 7. MSCs exposed to MesenPure™ require fewer cells to demonstrate robust differentiation

(A) BM-derived MSCs initially cultured in complete MesenCult™ medium without (Control) or with MesenPure™ were plated at three different densities and then differentiated using MesenCult™ Osteogenic Stimulatory Kit (Mouse, Catalog #05504). Differentiation cultures were fixed and stained for Alkaline Phosphatase (ALP) activity (red areas) and mineralization (black areas). In control cultures, modest differentiation into the osteogenic lineage was only observed when cultures were seeded at the highest density. In MesenPure™-exposed cultures, robust osteogenic differentiation was observed at all seeding densities. (B) 4.0 x 10 4 cells/cm 2 of P2 CB-derived MSCs were used in osteogenic and adipogenic differentiation. 5.0 x 10 5 cells of P1 CB-derived MSCs were used for chondrogenic differentiation. MesenPure™-exposed cultures displayed strong osteogenic, adipogenic and chondrogenic differentiation.

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.
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