Isolation of Mesenchymal Stem CellsExpansion of Human Mesenchymal Stem CellsDifferentiation of Human Mesenchymal Stem CellsFrequently Asked Questions

Mesenchymal stem cells (MSCs) have the capacity for multi-lineage differentiation, giving rise to a variety of mesenchymal phenotypes such as osteoblasts (bone), adipocytes (fat), and chondrocytes (cartilage). Stem cell therapy holds immense promise of delivering the next generation of future medical breakthroughs. In this respect, multipotent progenitor cells, such as hMSCs, have attracted high clinical interest because of their ability to differentiate into various cell types and their immunoregulatory properties. Together, these features enable the allogeneic use of hMSCs and thus make them an attractive target for commercial therapeutic development. Below you will find step-by-step protocols used to isolate, expand and differentiate MSCs properly in stem cell cultures.

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Mesenchymal stem cells have been isolated from a variety of tissues including human bone marrow, adipose tissue, umbilical cord and dental pulp. Below is a simple general protocol that can be used to derive MSCs from a variety of tissue sources.

Note: MSC populations will vary from donor to donor and might require further optimization.

Tissue culture plastic- or glassware plates should be coated with 0.1% gelatin as follows:

IMPORTANT: Do not vortex the cells.

IMPORTANT: Do not add the entire volume of media all at once to the cells. This may result in decreased cell viability due to osmotic shock.

IMPORTANT: Do not vortex the cells.

IMPORTANT: Seeding density should be 5,000-6,000 cells/cm2

NOTE: Depending on seeding density and passage number (i.e. later passages), cells may take longer to reach 80% confluency.

IMPORTANT: Subculture cells before reaching 100% confluency.

IMPORTANT: Do not vortex the cells.

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Mesenchymal Stem Cell Culture Protocols - Sigma-Aldrich

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