Abstract
BackgroundMesenchymal stromal cells (MSCs) are widely used in clinical trials for bone repair and regeneration. Despite previous evidence showing a prominent osteogenic potential of 2D cultured CD271 enriched MSCs, the osteogenic potential of CD271 enriched cells cultured on 3D scaffold is unknown. Apatite-wollastonite glass ceramic (A-W) is an osteoconductive biomaterial shown to be compatible with MSCs. This is the first study comparing the attachment, growth kinetics, and osteogenic potential of two MSC populations, namely heterogeneous plastic adherence MSCs (PA-MSCs) and CD271-enriched MSCs (CD271-MSCs), when cultured on A-W 3D scaffold.ResultsThe paired MSC populations were assessed for their attachment, growth kinetics and ALP activity using confocal and scanning electron microscopy and the quantifications of DNA contents and p-nitrophenyl (pNP) production respectively. While the PA-MSCs and CD271-MSCs had similar expansion and tri-lineage differentiation capacity during standard 2D culture, they showed different proliferation kinetics when seeded on the A-W scaffolds. PA-MSCs displayed a well-spread attachment with more elongated morphology compared to CD271-MSCs, signifying a different level of interaction between the cell populations and the scaffold surface. Following scaffold seeding PA-MSCs fully integrated into the scaffold surface and showed a stronger propensity for osteogenic differentiation as indicated by higher ALP activity than CD271-MSCs. Furthermore, A-W scaffold seeded uncultured non-enriched bone marrow mononuclear cells also demonstrated a higher proliferation rate and greater ALP activity compared to their CD271-enriched counterpart.ConclusionsOur findings suggest that CD271-positive enrichment of a population is not beneficial for osteogenesis when the cells are seeded on A-W scaffold. Furthermore, unselected heterogeneous MSCs or BM-MNCs are more promising for A-W scaffold based bone regeneration. This leads to a conclusion of broader clinical relevance for tissue engineering: on the basis of our observations here the osteogenic potential observed in 2D cell culture should not be considered indicative of likely performance in a 3D scaffold based system, even when one of the cell populations is effectively a subset of the other.
Highlights
Mesenchymal stromal cells (MSCs) are widely used in clinical trials for bone repair and regeneration
plastic adherence MSCs (PA-MSCs) and CD271-MSC show similar characteristics in 2D culture Paired MSCs were expanded in 2D culture up to 6 passages
The results demonstrated that both PA-MSCs and CD271-MSCs displayed the standard MSC characteristics including plastic adherent, fibroblast-like morphology (Fig. 2b, c), ability to differentiate into osteogenic, chondrogenic and adipogenic lineages (Fig. 2d, e), positive
Summary
Mesenchymal stromal cells (MSCs) are widely used in clinical trials for bone repair and regeneration. Mesenchymal stromal cells (MSCs) and bioactive scaffolds are increasingly used in translational research and clinical applications to enhance bone repair/regeneration [1, 2]. According to the minimal criteria to define human MSCs proposed by the International Society for Cellular Therapy (ISCT), among other features, MSCs should express surface markers CD73, CD90 and CD105 and lack of expression of haematopoietic markers CD45, CD34, CD19 and CD14 [5]. These minimal criteria were updated for adipose tissue derived MSCs as they express CD34 [6]. No study has been reported comparing heterogeneous and CD271-enriched MSC populations when cultured on 3D scaffold
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