Mesenchymal stem cells

The CD271 MicroBead Kit,  human was developed for the positive selection or depletion of cells expressing CD271 (LNGFR). They are ideally suited for the isolation of CD271⁺ MSCs from human tissues, including bone marrow and lipoaspirate, but have also been successfully used to isolate MSCs from bone marrow of monkey, goat, dog, pig, and sheep (PMID: 20367498).

The Anti-MSCA-1 (W8B2) MicroBead Kit,  human is based on the antibody clone W8B2, which recognizes the mesenchymal stromal cell antigen-1 (MSCA-1), a highly specific marker to isolate MSCs with a high proliferative potential from bone marrow .

Large-scale isolation and expansion of MSCs from human bone marrow is time and resource intensive. Success of this procedure depends on the quality of the starting material, and thus it is important to quantify the MSC content of the bone marrow. The number of MSCs in human blood source material is commonly determined based on the colony-forming–unit fibroblast (CFU-F) assay. However, the underlying process is time consuming and susceptible to serum lot variation, plating densities, and the subjective definition and scoring of colonies. It was shown that the Anti-MSCA-1 (W8B2) antibody exclusively detects CD271 (LNGFR)^bright cells, which are the only cells that give rise to CFU-F (PMID: 17395729). Moreover, there is a close linear relationship between the number of CFU-F colonies counted manually after 14 days of culture and the number of CD271 (LNGFR)^bright cells per milliliter of bone marrow aspirate (PMID: 22268519). Therefore, CD271 and MSCA-1 are valuable markers for the enumeration of MSCs by flow cytometry.

The BM-MSC Analysis Cocktail Kit, anti-human is designed for the quantification of MSCs in  human bone marrow samples, based on the expression of CD271 (LNGFR) and MSCA-1 (W8B2). The kit allows identification of CD45⁺ leukocytes and CD271 (LNGFR)⁺MSCA-1 (W8B2)⁺ MSCs that are also CD45^dim. The BM-MSC Analysis Cocktail Kit, anti-human standardizes the identification and quantification of human MSCs from fresh bone marrow aspirate by flow cytometry and includes details on the appropriate gating strategy.

A scientific poster describing the standardized quantification of MSCs by flow cytometry can be downloaded from the related resources panel to the right.

Miltenyi Biotec offers a streamlined and standardized method that enables the fast, easy, and reliable identification and phenotyping of  cultured MSCs according to standards defined by the International Society for Cellular Therapy (ISCT; PMID: 16923606). The MSC Phenotyping Cocktail Kit, anti-human, REAfinity includes all reagents necessary for MSC identification based on the expression of CD73, CD90, and CD105, and the absence of CD14, CD20, CD34, and CD45. The data sheet included in the kit provides details on the appropriate gating strategy. 

First protocols to isolate and expand MSCs from human tissues were based on media containing fetal calf serum (FCS). To meet increasing safety requirements from regulatory authorities, Miltenyi Biotec provides FCS-containing medium (StemMACS MSC Expansion Media), serum- and xeno-free medium (StemMACS MSC Expansion Media Kit XF), as well as serum-free, GMP-grade medium (MSC-Brew GMP Medium). All media enable reliable and reproducible expansion of MSCs from bone marrow, adipose tissue, and umbilical cord.

StemMACS™ MSC Expansion Media Kit XF is perfect for the direct isolation and expansion of human MSCs from various sources, including bone marrow and lipoaspirate. Its xeno-free composition enables fast expansion while maintaining differentiation potential and immunomodulation ability. MSCs cultured using the StemMACS MSC Expansion Media Kit XF show a superior expansion rate compared to other standard MSC culture methods.

Differentiation potential is a meaningful feature to qualify and define an MSC population. The propensity of MSCs to differentiate into adipocytes, chondrocytes, or osteoblasts is influenced by MSC origin. MSCs from bone marrow, for example, show greater potential to differentiate into osteoblasts than MSCs from umbilical cord. Increased age of donor and passaging of cultured MSCs also decrease the differentiation potential.

StemMACS™ AdipoDiff, OsteoDiff, and ChondroDiff Media are optimized differentiation media for human MSCs from various tissue sources. The media support the analysis or quality control of the differentiation capacity of expanded MSCs. They also facilitate in vitro studies on the processes of MSC differentiation, including gene expression and protein profiling.

During the last few years, scientists have directed their attention towards the immunomodulatory potential of MSCs. It has been observed that MSCs derived from bone marrow suppress the proliferation of T cells (PMID: 11986244, 11823036). This function of MSCs can be analyzed using the MSC Suppression Inspector, human, which contains T cell stimulation reagents optimized for an MSC suppression assay.