Bone marrow- and adipose tissue-derived mesenchymal stem cells (MSCs)

  • MSC expansion in xeno- and serum-free medium
  • Optimized flow cytometry reagents for quantification and phenotyping analyses
  • Complete workflow from MSC isolation and expansion, to differentiation and immunomodulation assays 

Application protocols

Discover our different mesenchymal stem cell workflows and find the one that fits your experimental needs.

Application data by workflow step

Quantification of MSCs in bone marrow aspiration 

The MSC Enumeration Kit was developed for the standardized, quick, and absolute quantification of human MSCs from fresh bone marrow aspirate by flow cytometry.  It takes only 30 minutes and requires only 200 µl of bone marrow sample per test. 

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Enumeration of MCSs by flow cytometry

The MSC Enumeration Kit, human  provides a cocktail of 3 fluorochrome-conjugated antibodies that are used to identify MSCs, These include the MSC-specific marker CD271 (LNGFR) and exclusion markers CD45 and CD235a. Anti-MSCA-1 (W8B2) is provided separately in the kit and can be added to specifically identify MSCs with high proliferative potential.

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Scientific poster
Standarized quantification of human bone marrow mesenchymal stromal cells based on a flow cytometry assay

Kathrin Godthardt, Claudia Schreiner, Andreas Bosio, and Sebastian Knöbel, Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany

Enrichment of MSCs from bone marrow and adipose tissue

Historically, a number of methods relying on specific physical properties have been used to isolate MSCs from sites at which they reside. The problem with this type of approach is that no physical properties have been uniquely ascribed to MSCs. Therefore, many different cell types are co-isolated, resulting in a mixed population of cells.

Magnetic cell isolation (positive selection) of CD271 (LNGFR)+ cells results in a cell population containing an approximately 1000-fold greater concentration of MSCs as compared to conventional methods of isolation. This enables isolation of a homogeneous, and consequently more effective non-hematopoietic stem cell population.

Enrichment of CD271 (LNGFR)+ MSCs from bone marrow aspirate using the CD271 MicroBead Kit (APC), human
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Positive selection of CD271(LNGFR)+ or MSCA-1 (W8B2)+ cells

CD271 MicroBead Kits were developed for the isolation of CD271 (LNGFR)+ MSCs from bone marrow or adipose tissue with high yield and purity.

The Anti-MSCA-1 (W8B2) MicroBead Kit  was developed for the isolation of MSCA-1 (W8B2)+ MSCs with high proliferative capacity from bone marrow aspirates.


MSC expansion

Our cell culture options provide reliable solutions for the maintenance and expansion of MSCs with preserved differentiation potential and immunomodulation ability.

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StemMACS™ MSC Expansion Media Kit XF, human

  • Robust expansion of human MSCs in xeno- and serum-free conditions
  • No substrate pre-coating of the culture vessels required
  • No additional supplement or growth factors or serum required
  • Upgrade to MACS GMP medium possible to facilitate clinical translation


Phenotyping of in vitro expanded MSCs

The MSC Phenotyping Kit was developed for the standardized identification and phenotyping of cultured human MSCs by flow cytometry based on the defined ISCT standards.

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MSC Phenotyping Kit

Phenotyping based on the expression of CD73, CD90, CD105 and exclusion of CD14, CD20, CD34, and CD45 markers

Anibodies for isotype controls and compensation of PerCP, PE, APC, and FITC channels are included 


Differentiation of MSCs

We provide a variety of media options that support the differentiation of MSCs into adipocytes, osteoblasts, and chondrocytes. The different StemMACS Differentiation Media are suitable for the analysis or quality control of the differentiation capacity of expanded MSCs, as well as in vitro studies focusing on the processes involved in MSC differentiation, including gene expression and protein profiling.

Adipocytes stained with Oil Red O after cultivation of MSCs for 21 days in StemMACS AdipoDiff Medium.
Osteoblasts differentiated from human MSCs after cultivation for ten days in StemMACS OsteoDiff Medium stained with NBT substrate.
Chondrocytes stained for aggrecan (red) and nuclei (blue) after cultivation of MSCs for 21 days in StemMACS ChondroDiff Medium.

Characterization of MSC immunomodulatory function

The suppression potential of MSCs varies among in vitro expanded cells and shows donor-dependent differences. MSCs can be “licensed” by inflammatory cytokines such IFN-γ and TNF-α to become more immunosuppressive and show a more homogeneous phenotype in this regard. The MSC Suppression Inspector, human was developed for standardized characterization of the immunomodulatory function of MSCs by co-culture with CD4+CD25 or CD4+ responder T (Tresp) cells 

Evaluation of T cell-specific immunmodulatory function of  PA-MSCs or CD271+ MSCs with the MSC Suppression Inspector, human
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MSC Suppression Inspector, human

  • The kit contains Anti-Biotin MACSiBead Particles pre-loaded with biotinylated CD2, CD3, and CD28 antibodies for optimal T cell stimulation

  • Protocol includes Tresp isolation and stimulation, MSC-Tresp co-culture, and analysis of Tresp cell proliferation by 3H-thymidine incorporation, as well as carboxyfluorescein succinimidyl ester (CFSE) staining

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Application note
Effective licensing of human mesenchymal stem cells

This application note describes the licensing of MSCs using IFN-γ and TNF-α and a procedure to analyze marker expression and the immunosuppressive characteristics of MSCs using the MSC Suppression Inspector, human.

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