Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement

Dominici, Massimo; Blanc, Katarina Le; Mueller, Ingo; Slaper‐Cortenbach, Ineke; Marini, Frank C.; Krause, Diane S.; Deans, Robert; Keating, Armand; Prockop, Darwin J.; Horwitz, Edwin M.

doi:10.1080/14653240600855905

Cited by 15,138 publications

(13,400 citation statements)

References 8 publications

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“…2B). cGFP‐DPPCs were found to positively express mesenchymal stem cell markers CD73, CD90 and CD105, and were negative for the hematopoietic marker CD45 consistent with standard MSC characterisation 23, 24. In addition to the mesenchymal stem cell markers, these cells express CD146, indicative of pericytes but have additional expression of CD34, suggesting potential presence of transit amplifying progenitor cells 25.…”

Section: Resultssupporting

confidence: 55%

A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells

et al. 2015

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Harnessing mesenchymal stem cells for tissue repair underpins regenerative medicine. However, how the 3D tissue matrix maintains such cells in a quiescent state whilst at the same time primed to respond to tissue damage remains relatively unknown. Developing more physiologically relevant 3D models would allow us to better understand the matrix drivers and influence on cell‐lineage differentiation in situ. In this study, we have developed an ex vivo organotypic rat mandible slice model; a technically defined platform for the culture and characterization of dental pulp progenitor cells expressing GFP driven by the β‐actin promoter (cGFP DPPCs). Using confocal microscopy we have characterized how the native environment influences the progenitor cells transplanted into the dental pulp. Injected cGFP‐DPPCs were highly viable and furthermore differentially proliferated in unique regions of the mandible slice; in the dentine region, cGFP‐DPPCs showed a columnar morphology indicative of expansion and lineage differentiation. Hence, we demonstrated the systematic capacity for establishing a dental pulp cell‐micro‐community, phenotypically modified in the tooth (the “biology”); and at the same time addressed technical challenges enabling the mandible slice to be accessible on platforms for high‐content imaging (the biology in a “multiplex” format). © 2015 The Authors. Published by Wiley Periodicals, Inc.

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Section: Resultssupporting

confidence: 55%

A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells

et al. 2015

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“…According to the standards defined by the International Society for Cellular Therapy (ISCT), MSCs should be adherent to plastic under normal culturing conditions, have fibroblast-like morphology and show a specific immunophenotype measured by flow cytometry [23,24]. In concordance with these guidelines, AT-MSC donor cell lines grown in the FBS-supplemented medium expressed surface markers CD73, CD90 and CD105, and lacked the expression of CD14, CD19, CD45 and HLA-DR (Table 3).…”

Section: Resultsmentioning

confidence: 99%

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Kornilov

Puhka

Mannerström

et al. 2018

J of Extracellular Vesicle

156

125

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Fetal bovine serum (FBS) is the most commonly used supplement in studies involving cell-culture experiments. However, FBS contains large numbers of bovine extracellular vesicles (EVs), which hamper the analyses of secreted EVs from the cell type of preference and, thus, also the downstream analyses. Therefore, a prior elimination of EVs from FBS is crucial. However, the current methods of EV depletion by ultracentrifugation are cumbersome and the commercial alternatives expensive. In this study, our aim was to develop a protocol to completely deplete EVs from FBS, which may have wide applicability in cell-culture applications. We investigated different EV-depleted FBS prepared by our novel ultrafiltration-based protocol, by conventionally used overnight ultracentrifugation, or commercially available depleted FBS, and compared them with regular FBS. All sera were characterized by nanoparticle tracking analysis, electron microscopy, Western blotting and RNA quantification. Next, adipose-tissue mesenchymal stem cells (AT-MSCs) and cancer cells were grown in the media supplemented with the three different EV-depleted FBS and compared with cells grown in regular FBS media to assess the effects on cell proliferation, stress, differentiation and EV production. The novel ultrafiltration-based protocol depleted EVs from FBS clearly more efficiently than ultracentrifugation and commercial methods. Cell proliferation, stress, differentiation and EV production of AT-MSCs and cancer cell lines were similarly maintained in all three EV-depleted FBS media up to 96 h. In summary, our ultrafiltration protocol efficiently depletes EVs, is easy to use and maintains cell growth and metabolism. Since the method is also cost-effective and easy to standardize, it could be used in a wide range of cell-culture applications helping to increase comparability of EV research results between laboratories.

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“…We adhere herein, in concordance with the ISCT position paper of 2006 37, to the term mesenchymal stromal cell (MSC) rather mesenchymal stem cell since it is rarely that authors have demonstrated the stem‐like qualities, of self‐renewal and multilineage differentiation potential of offspring, of the cells they have isolated from the human umbilical cord. One exception to this was the work reported by Sarugaser et al 38 that did show, at the single cell clonal level, that true self‐renewing cells existed in the perivascular tissue of the cord that gave rise to multilineage offspring—work that was only possible due to the high CFU‐F frequency in the cell populations extracted from the perivascular tissue (see Fig.…”

Section: Wharton's Jelly As a Source Of Mscsmentioning

confidence: 87%

“…Indeed, the first of the minimal criteria set out by the ISCT 37 is that the cells are culture adherent. Taken literally, this would mean that MSC can only be described ex vivo; yet conventional wisdom holds that such cells—both mesenchymal cells and mesenchymal stem cells, do exist in vivo.…”

Section: Cell Phenotypementioning

confidence: 99%

Concise Review: Wharton's Jelly: The Rich, but Enigmatic, Source of Mesenchymal Stromal Cells

Davies

Walker

Keating

2017

Stem Cells Translational Medicine

162

145

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The umbilical cord has become an increasingly used source of mesenchymal stromal cells for preclinical and, more recently, clinical studies. Despite the increased activity, several aspects of this cell population have been under‐appreciated. Key issues are that consensus on the anatomical structures within the cord is lacking, and potentially different populations are identified as arising from a single source. To help address these points, we propose a histologically based nomenclature for cord structures and provide an analysis of their developmental origins and composition. Methods of cell isolation from Wharton's jelly are discussed and the immunophenotypic and clonal characteristics of the cells are evaluated. The perivascular origin of the cells is also addressed. Finally, clinical trials with umbilical cord cells are briefly reviewed. Interpreting the outcomes of the many clinical studies that have been undertaken with mesenchymal stromal cells from different tissue sources has been challenging, for many reasons. It is, therefore, particularly important that as umbilical cord cells are increasingly deployed therapeutically, we strive to better understand the derivation and functional characteristics of the cells from this important tissue source. Stem Cells Translational Medicine 2017;6:1620–1630

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Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement

Cited by 15,138 publications

References 8 publications

A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells

A 3D ex vivo mandible slice system for longitudinal culturing of transplanted dental pulp progenitor cells

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Concise Review: Wharton's Jelly: The Rich, but Enigmatic, Source of Mesenchymal Stromal Cells

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