The Myelopoietic Supportive Capacity of Mesenchymal Stromal Cells Is Uncoupled from Multipotency and Is Influenced by Lineage Determination and Interference with Glycosylation

Abstract

Cultured bone marrow stromal cells create an in vitro milieu supportive of long-term hemopoiesis and serve as a source for multipotent mesenchymal progenitor cells defined by their ability to differentiate into a variety of mesodermal derivatives. This study aims to examine whether the capacity to support myelopoiesis is coupled with the multipotency. Our results show that the myelopoietic supportive ability of stromal cells, whether from the bone marrow or from embryo origin, is not linked with multipotency; cell populations that possess multipotent capacity may or may not support myelopoiesis, whereas others, lacking multipotency, may possess full myelopoietic supportive ability. However, upon differentiation, the ability of multipotent mesenchymal progenitors to support myelopoiesis is varied. Osteogenic differentiation did not affect myelopoietic supportive capacity, whereas adipogenesis resulted in reduced ability to support the maintenance of myeloid progenitor cells. These differences were accompanied by a divergence in glycosylation patterns, as measured by binding to lectin microarrays; osteogenic differentiation was associated with an increased level of antennarity of N-linked glycans, whereas adipogenic differentiation caused a decrease in antennarity. Inhibition of glycosylation prior to seeding the stroma with bone marrow cells resulted in reduced capacity of the stromal cells to support the formation of cobblestone areas. Our data show that myelopoietic support is unrelated to the multipotent phenotype of cultured mesenchymal progenitors but is dependent on the choice of differentiation pathway and upon correct glycosylation of the stromal cells.

N-Glycosylation Profile of Undifferentiated and Adipogenically Differentiated Human Bone Marrow Mesenchymal Stem Cells: Towards a Next Generation of Stem Cell Markers

“…A significant increase of biantennary fucosylated N-glycans H3N4F1, H5N4F3, and S2H5N4F1 was observed in 15 days adipogenically differentiated MSCs, whereas undifferentiated human MSCs contained significantly more triantennary H6N5F1 and S1H6N5 and tetraantennary H7N6F1, S1H7N6, and S1H7N6F1 N-glycans than adipogenically differentiated MSCs. Our results are corroborated by Morad et al [44], who showed using lectin staining of membrane extracts that adipogenically differentiation of MSCs was accompanied by a decreased level of antennarity of complex-type N-glycans, namely of tri-and tetraantennary structures. In addition, they proved that the level of these structures was increased in osteogenically differentiated MSCs when compared to undifferentiated MSCs.…”

“…In addition, they proved that the level of these structures was increased in osteogenically differentiated MSCs when compared to undifferentiated MSCs. However, in the latter mentioned study, glycoanalysis of membrane proteins was performed using lectin microarrays, showing that different methods lead to similar results [44]. In contrast, Heiskanen et al reported that the acidic N-glycan profile of MSCs contained relatively more of the larger complex-type N-glycans (five GlcNAc or more) and more Nglycans with two or more fucose residues when compared to their osteogenically counterparts [37].…”

Bone Marrow Regeneration Promoted by Biophysically Sorted Osteoprogenitors From Mesenchymal Stromal Cells

“…Part of this shift reflects greater appreciation that MSC populations are heterogeneous, particularly when culture-expanded to large numbers in vitro, and include a subset of mesenchymal stem cells that exhibit in vitro self-renewal capacity or multilineage differentiation [8,9]. First is the implication that MSC potency may also vary significantly with the tissue or microenvironment of origin [10]. Several studies have established that MSCs derived from adipose tissue, umbilical cord, and BM have different gene expression profiles, although MSCs from each source can exhibit multipotent differentiation [11,12].…”

À la recherche d’une définition moléculaire plus que descriptive pour les cellules souches

“…Toutefois certaines propriétés des CSM ne résultent pas d'un processus de différenciation au sens strict du terme : c'est le cas du rôle de support de l'hématopoïèse qu'exercent certaines cellules stromales mésenchymateuses. De fait, les CSM indifférenciées, mais aussi les précurseurs adipocytaires, possèdent aussi cette propriété [7]. Si cette capacité de soutien est perdue lorsque ces précurseurs se différencient en adipocytes matures accumulant des lipides, tel n'est pas le cas lors de la différenciation des CSM en précurseurs ostéogéniques puis en ostéocytes sécrétant une matrice osseuse [7].…”

“…De fait, les CSM indifférenciées, mais aussi les précurseurs adipocytaires, possèdent aussi cette propriété [7]. Si cette capacité de soutien est perdue lorsque ces précurseurs se différencient en adipocytes matures accumulant des lipides, tel n'est pas le cas lors de la différenciation des CSM en précurseurs ostéogéniques puis en ostéocytes sécrétant une matrice osseuse [7]. Cette organisation hiérarchique de cellules qui perdent progressivement leur potentiel au fur et à mesure qu'elles s'éloignent du progéniteur ancestral ne semble donc pas s'appliquer au processus de différenciation des CSM.…”