Abstract
Osteoarthritis is characterized by loss of articular cartilage also due to reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients. Adipose tissue is an attractive source of MSCs (ATD-MSCs), representing an effective tool for reparative medicine, particularly for treatment of osteoarthritis, due to their chondrogenic and osteogenic differentiation capability. The treatment of symptomatic knee arthritis with ATD-MSCs proved effective with a single infusion, but multiple infusions could be also more efficacious. Here we studied some crucial aspects of adipose tissue banking procedures, evaluating ATD-MSCs viability, and differentiation capability after cryopreservation, to guarantee the quality of the tissue for multiple infusions. We reported that the presence of local anesthetic during lipoaspiration negatively affects cell viability of cryopreserved adipose tissue and cell growth of ATD-MSCs in culture. We observed that DMSO guarantees a faster growth of ATD-MSCs in culture than trehalose. At last, ATD-MSCs derived from fresh and cryopreserved samples at −80°C and −196°C showed viability and differentiation ability comparable to fresh samples. These data indicate that cryopreservation of adipose tissue at −80°C and −196°C is equivalent and preserves the content of ATD-MSCs in Stromal Vascular Fraction (SVF), guaranteeing the differentiation ability of ATD-MSCs.
Highlights
In the last years many studies have been focused on tissue regeneration and reparative medicine, to cure osteoarthritis, characterized by loss of articular cartilage due to a reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients [1]
Adipose tissues harvested with (n 35)/without (n 23) local anesthetic maintained normal histological structure without evident differences between 2 groups. Both with and without anesthetic, the tissues were compact, with polygonal cells and good stroma, without any evidence of fatty tissue degeneration or necrosis (Figures 1(a) and 1(b)). These data were confirmed by the immunohistochemical analysis (IHC) analysis, showing that the different cellular components of adipose tissue, stained with specific antibodies, resulted comparably in all analysed samples (Figures 1(c)–1(f))
Regenerative and in particular reparative medicine represent the new frontier for orthopaedic diseases, such as osteoarthritis, that can benefit of treatments with ATD-MSCs
Summary
In the last years many studies have been focused on tissue regeneration and reparative medicine, to cure osteoarthritis, characterized by loss of articular cartilage due to a reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients [1]. Adult MSCs, originally retrieved from bone marrow [2], can be isolated from adipose tissue with higher frequency than in bone marrow [3] and show a great seductive potential to the orthopaedic community for their reparative capabilities and differentiation ability towards adipogenic, osteogenic, and chondrogenic lineages [4, 5], to home in injured tissues, to release factors for wound healing, and to modulate the immune system [5,6,7]. ATD-MSCs express many markers common to bone marrow-derived stem cells [10, 11], such as CD90, CD44, CD73, CD105, and CD271 [5, 12]. CD271 has been proposed as marker of primary choice for tissue regeneration by ATD-MSCs in older subjects, since this subpopulation is maintained in elderly people [13], and CD271+ MSCs are associated with a high efficiency of proliferation and trilineage differentiation compared to CD271− MSCs [12, 14].
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