Enrichment of CD146⁺ Adipose-Derived Stem Cells in Combination with Articular Cartilage Extracellular Matrix Scaffold Promotes Cartilage Regeneration

Abstract

Heterogeneity of mesenchymal stem cells (MSCs) influences the cell therapy outcome and the application in tissue engineering. Also, the application of subpopulations of MSCs in cartilage regeneration remains poorly characterized. CD146+ MSCs are identified as the natural ancestors of MSCs and the expression of CD146 are indicative of greater pluripotency and self-renewal potential. Here, we sorted a CD146 + subpopulation from adipose-derived mesenchymal stem cells (ADSCs) for cartilage regeneration. Methods : CD146 + ADSCs were sorted using magnetic activated cell sorting (MACS). Cell surface markers, viability, apoptosis and proliferation were evaluated in vitro . The molecular signatures were analyzed by mRNA and protein expression profiling. By intra-articular injections of cells in a rat osteochondral defect model, we assessed the role of the specific subpopulation in cartilage microenvironment. Finally, CD146 + ADSCs were combined with articular cartilage extracellular matrix (ACECM) scaffold for long term (3, 6 months) cartilage repair. Results : The enriched CD146 + ADSCs showed a high expression of stem cell and pericyte markers, good viability, and immune characteristics to avoid allogeneic rejection. Gene and protein expression profiles revealed that the CD146 + ADSCs had different cellular functions especially in regulation inflammation. In a rat model, CD146 + ADSCs showed a better inflammation-modulating property in the early stage of intra-articular injections. Importantly, CD146 + ADSCs exhibited good biocompatibility with the ACECM scaffold and the CD146 + cell-scaffold composites produced less subcutaneous inflammation. The combination of CD146 + ADSCs with ACECM scaffold can promote better cartilage regeneration in the long term. Conclusion : Our data elucidated the function of the CD146 + ADSC subpopulation, established their role in promoting cartilage repair, and highlighted the significance of cell subpopulations as a novel therapeutic for cartilage regeneration.

Evolution of ASC Immunophenotypical Subsets During Expansion In Vitro

“…Similarly, it is intriguing that a subset of these cells exhibiting the CD31−CD34+CD146− profile, previously found to identify a stem cell lineage [25,64,65], also grew proportionally to the culture period. In addition, possibly important ramifications may reveal that some antigens that have been ambiguously associated with stemness, including CD200, CD248, and CD271 [66,67], were reinforced during the culture, while all subsets featuring CD146, which has previously been recognized as a surrogate for stemness [40,[68][69][70][71] were sequestered. Concerning interpersonal variability, the spread of the data roughly follows previously reported trends [49,72], except for two specific subsets, the CD166+CD271−CD248+ and CD166+CD271+CD248+, which were identified only in a single donor.…”

Autologous Fractionated Adipose Tissue as a Natural Biomaterial and Novel One-Step Stem Cell Therapy for Repairing Articular Cartilage Defects

“…Furthermore, fragmented fat tissue as a natural biomaterial for efficient drug delivery, improved the local drug concentration and prolonged its therapeutic activity ( Alessandri et al, 2019 ), indicating that our ECM/SCF-gel may act as a self-controlled drug release system of intrinsic growth factors to promote cartilage repair. Different from scaffold-based ADSCs treatment ( Li X. et al, 2019 ), this strategy allows overcoming the limitations associated with the enzymatic isolation of SVF cells from adipose tissue and the expansion of ADSCs, as well as the complex process of scaffold fabrication. In addition, a recent study reported that after cryopreservation at −20°C for 3 months without a cryoprotectant, the mechanically processed fat product SVF gel from human lipoaspirate maintained ECM integrity and ADSC viability as well as their multipotency ( Feng et al, 2019 ).…”

Adipose Tissue-Derived Stem Cells: The Biologic Basis and Future Directions for Tissue Engineering

“…The immunophenotype of ASCs can express other essential factors involved in stemness, self-renewal and differentiation potentials, such as CD146 and CD271. They are also related to enhanced capacity for healing bone defects or cartilage or to differential paracrine wound healing activity [ 40 , 41 ].…”

“…Li and collaborators isolated a CD146 + subpopulation from ASCs and tested for cartilage regeneration [ 40 ]. Cartilage lesions typically result in inflammation and represent an important test in cartilage repair [ 42 ].…”