Ex vivo identification and characterization of a population of CD13high CD105+ CD45− mesenchymal stem cells in human bone marrow

Abstract

IntroductionMesenchymal stem cells (MSCs) are multipotent cells capable of self-renewal and multilineage differentiation. Their multipotential capacity and immunomodulatory properties have led to an increasing interest in their biological properties and therapeutic applications. Currently, the definition of MSCs relies on a combination of phenotypic, morphological and functional characteristics which are typically evaluated upon in vitro expansion, a process that may ultimately lead to modulation of the immunophenotypic, functional and/or genetic features of these cells. Therefore, at present there is great interest in providing markers and phenotypes for direct in vivo and ex vivo identification and isolation of MSCs.MethodsMultiparameter flow cytometry immunophenotypic studies were performed on 65 bone marrow (BM) samples for characterization of CD13high CD105+ CD45– cells. Isolation and expansion of these cells was performed in a subset of samples in parallel to the expansion of MSCs from mononuclear cells following currently established procedures. The protein expression profile of these cells was further assessed on (paired) primary and in vitro expanded BM MSCs, and their adipogenic, chondrogenic and osteogenic differentiation potential was also determined.ResultsOur results show that the CD13high CD105+ CD45− immunophenotype defines a minor subset of cells that are systematically present ex vivo in normal/reactive BM (n = 65) and that display immunophenotypic features, plastic adherence ability, and osteogenic, adipogenic and chondrogenic differentiation capacities fully compatible with those of MSCs. In addition, we also show that in vitro expansion of these cells modulates their immunophenotypic characteristics, including changes in the expression of markers currently used for the definition of MSCs, such as CD105, CD146 and HLA-DR.ConclusionsBM MSCs can be identified ex vivo in normal/reactive BM, based on a robust CD13high CD105+ and CD45− immunophenotypic profile. Furthermore, in vitro expansion of these cells is associated with significant changes in the immunophenotypic profile of MSCs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0152-8) contains supplementary material, which is available to authorized users.

KIT D816V–mutated bone marrow mesenchymal stem cells in indolent systemic mastocytosis are associated with disease progression

“…39 These results are in line with recent observations by Nemeth et al who described the presence of abnormal MSCs with slow proliferation, signs of senescence, and impaired osteogenic function (vs normal MSC colonies) in SM patients. 60 Of note, these authors did not find the KIT D816V mutation in cultured BM MSCs from any of the 5 SM patients they analyzed.…”

“…2 BM cells showed immunophenotypic features which were fully consistent with previously defined criteria for MSCs [39][40][41] such as: absence of CD11b, CD14, CD19, CD34, and CD45 expression; heterogeneous reactivity for CD10 and HLA-DR; and expression of the CD44, CD73, CD90, CD105, CD140b, CD146, CD271, MSCA-1, SSEA-4, and STRO-1 MSC-associated markers (supplemental Figure 1, available on the Blood Web site).…”

Phenotypic Characterization of Bone Marrow Mononuclear Cells and Derived Stromal Cell Populations from Human Iliac Crest, Vertebral Body and Femoral Head

“…Rasini et al, 2013 reported site-specific phenotypes based on cell-surface marker expression between cells in different anatomical localizations [16]. Previous studies focused on characterizing individual BMSC populations in human bone marrow within the freshly isolated MNC fraction: CD45 low /D7FIB+/CD271+ [11]; lin−/CD271+/CD45−/CD146+ [17]; CD64 bright /CD31 bright /CD14 neg [18]; CD13 high /CD105+/CD45− [19]; CD145−/CD34−/CD146+ [20,21,22]; PDPN+/CD146−CD73+CD164+ [23]. Whether the abundance of these discrete BMSC subsets may determine the yield and/or the phenotypic characteristics of the subsequently in vitro derived stromal population remains unclear.…”

Bone Tissue Engineering Using Human Cells: A Comprehensive Review on Recent Trends, Current Prospects, and Recommendations

“…These surface markers are used to purify BM-MSCs by fluorescence-activated cell sorting (FACS) as well as the magnetic-activated cell sorting (MACS). In addition, the use of Percoll gradient centrifugation and adherence to tissue-culture treated plastics are proposed as more simple techniques to purify BM-MSCs [163]. BM-MSCs can be used as undifferentiated or fully differentiated cells for TE applications.…”