Isolation and Characterization of Human Mesenchymal Stromal Cell Subpopulations: Comparison of Bone Marrow and Adipose Tissue

Abstract

Preparations of mesenchymal stromal cells (MSCs) are generally obtained from unfractionated tissue cells, resulting in heterogeneous cell mixtures. Several markers were proposed to enrich these cells, but the majority of these markers are defined for bone marrow (BM). Moreover, the surface markers of freshly isolated MSCs also differ from those of cultured MSCs in addition to a phenotypic variation depending on the MSC source. For tissue engineering applications, it is crucial to start with a well-defined cell population. In this study, we performed immunomagnetic selections with five single surface markers to isolate MSC subpopulations from BM and adipose tissue (AT): CD271, SUSD2, MSCA-1, CD44, and CD34. We determined the phenotype, the clonogenicity, the proliferation, the differentiation capacity, and the immunoregulatory profile of the subpopulations obtained in comparison with unselected cells. We showed that native BM-MSCs can be enriched from the positive fractions of MSCA-1, SUSD2, and CD271 selections. In contrast, we observed that SUSD2 and MSCA-1 were unable to identify MSCs from AT, meaning they are not expressed in situ. Only the CD34(+) selection successfully isolated MSCs from AT. Interestingly, we observed that CD271 selection can define AT cell subsets with particular abilities, but only in lipoaspiration samples and not in abdominoplasty samples. Importantly, we found a population of clear CD34(+) fresh BM-MSCs displaying different properties. A single marker-based selection for MSC enrichment should be more advantageous for cell therapy and would enable the standardization of efficient and safe therapeutic intervention through the use of a well-identified and homogeneous cell population.

Isolation and characterization of primary bone marrow mesenchymal stromal cells

“…RECs exhibited robust multilineage differentiation and selfrenewal potency, and were therefore considered to be the most primitive and undifferentiated population. 49 Last, in addition to CD146 and THY-1, other markers, such as MSCA-1 and SUSD2, have also been used to isolate subpopulations of MSCs, 50 and furthermore, differences in differentiation potential of single-sorted BMSC-derived CFU-Fs point to a possible hierarchical structure of the primary MSC compartment. 15 Here, our current ongoing single-cell gene expression experiments will hopefully contribute to further clarify this important aspect of primary MSC biology.…”

Studies of the Influence of Gold Nanoparticles on Characteristics of Mesenchymal Stem Cells

“…This minimum set of phenotypic criteria for MSCs identification is not associated with their origin [14]. MSCs are normally taken from various tissues, such as bone marrow, fat, and muscles involved in tissue homeostasis and regeneration [15]. We selected bone marrow MSCs of rats as a model object for studying the effect of AuNPs on stem cells.…”

Extracellular matrix and α5β1 integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells

“…Staining of human adipose tissue for α 5 -integrin identified a population of perivascular cells, and sorting of α 5 -integrin-positive SVF cells separated all clonogenic progenitors. SVF cells are highly heterogeneous and numerous markers have been proposed so far to specifically isolate clonogenic, osteogenic progenitors444546. Our data show that α 5 -integrin, in combination with CD73, could be used to enrich for clonogenic progenitors but is not sufficiently specific for their selective isolation from SVF cells.…”