Contaminating hematopoietic cells are a common problem when isolating and expanding mouse mesenchymal stem and progenitors cells (MSCs). Both MSCs and hematopoietic cells are plastic adherent and form colonies. However, they can be distinguished by their size and appearance. MSCs appear as fibroblast-like cells that are bigger, flatter and less refractive than hematopoietic cells when visualized using phase contrast light microscopy. When culturing with traditional medium, it can take multiple passages to reduce the hematopoietic cell population to achieve an enriched mouse MSC population. This can lead to senescence and a decrease in the functionality of mouse MSCs with each subsequent passage.
MesenCult™ Expansion Kit (Mouse: Catalog #05513)
contains medium and reagents to derive and culture-expand mouse MSCs from bone marrow, compact bone, adipose tissue and mouse embryonic fibroblasts (MEFs). This kit contains MesenPure™, a proprietary supplement, that when added to complete MesenCult™ Expansion medium can enrich for mouse MSCs as early as passage 0.
These are considerations when deriving and expanding mouse MSCs:
- Culture mouse MSCs under hypoxic conditions to promote faster expansion. It is shown that mouse MSCs expand faster under hypoxic conditions (5% O2) than under normoxic conditions (20% O2).1 Our data also show increased mouse MSC expansion under hypoxic conditions (Figure 1), which can be achieved by using either a hypoxic incubator or a hypoxic chamber (
- Add MesenPure™, which is included in the MesenCult™ Expansion Kit (Mouse; Catalog #05513), to complete MesenCult™ Expansion medium to enrich for mouse MSCs as early as passage 0. MesenPure™ reduces hematopoietic cells and promotes faster expansion of MSCs. As a result, fewer passages are required to obtain an enriched population of mouse MSCs (Figure 1, Table 1).
- Isolate mouse MSCs from compact bone. A video protocol to harvest mouse MSCs from compact bone is available:
Isolation of MSCs from Mouse Compact Bone
. When culturing mouse MSCs with MesenPure™ under hypoxic conditions, higher numbers of CFU-F colonies are observed in compact bone (140 ± 20 x 106 cells; n=3) than in bone marrow (109 ± 11 x 106 cells; n=3).
For more information on deriving and expanding mouse MSCs, please refer to the MesenCult™ Expansion Kit for mouse MSCs’ Product Information Sheet. If you have any questions about this product, or about other media and reagents available for MSC research, please contact firstname.lastname@example.org.