Introduction. The Infrapatellar fat pad (IPFP) represents an emerging alternative source of adipose-derived mesenchymal stem cells (ASCs). We compared the characteristics and differentiation capacity of ASCs isolated from IPFP and SC. Materials and Methods. ASCs were harvested from either IPFP or SC. IPFPs were collected from patients undergoing total knee arthroplasty (TKA), whereas subcutaneous tissues were collected from patients undergoing lipoaspiration. Immunophenotypes of surface antigens were evaluated. Their ability to form colony-forming units (CFUs) and their differentiation potential were determined. The ASCs karyotype was evaluated. Results. There was no difference in the number of CFUs and size of CFUs between IPFP and SC sources. ASCs isolated from both sources had a normal karyotype. The mesenchymal stem cells (MSCs) markers on flow cytometry was equivalent. IPFP-ASCs demonstrated significantly higher expression of SOX-9 and RUNX-2 over ASCs isolated from SC (6.19 ± 5.56-, 0.47 ± 0.62-fold; p value = 0.047, and 17.33 ± 10.80-, 1.56 ± 1.31-fold; p value = 0.030, resp.). Discussion and Conclusion. CFU assay of IPFP-ASCs and SC-ASCs harvested by lipoaspiration technique was equivalent. The expression of key chondrogenic and osteogenic genes was increased in cells isolated from IPFP. IPFP should be considered a high quality alternative source of ASCs.
Objectives The objective of this study was to investigate iron sucrose labeling in mesenchymal stem cell (MSCs) tracking. Background Adipose-derived mesenchymal stem cell-based therapy is a promising strategy for promoting musculoskeletal repair. Methods Iron sucrose-labeled adipose-derived mesenchymal stem cells (IS-labeled ASCs) were tracked using T2-and T2∗-weighted sequences by 1.5 and 3 T MRI in an in vitro model. ASCs were isolated from cosmetic liposuction specimens. ASCs from passages 4–6 were labeled with iron sucrose (Venofer®) which was added to the cell culture medium. Pre- and post-iron sucrose labeled ASCs were evaluated for cell surface immunophenotypes. Cell viability as well as chondrogenic, adipogenic and osteogenic differentiation of IS-labeled-ASCs were evaluated. The IS-labeled ASCs were titrated into microtubes at 1 × 10 3 , 1 × 10 4 , 1 × 10 5 and 1 × 10 6 cells/ml/microtube and their intensities were determined by 1.5 and 3T MRI using T2-and T2∗-weighted sequences. Results The expression markers of IS-labeled ASCs from flow cytometry were equivalent to control. The mean cell viability was 97.73 ± 2.06%. Cell differentiations of IS-labeled ASCs were confirmed in each lineage using specific staining solutions. T2∗-weighted sequences (T2∗) were able to detect iron sucrose labeled-ASCs at a minimum of 1 × 10 5 cells/ml/microtube using 1.5 and 3T MRI, but the detection sensitivity was lower with T2-weighted sequences (T2). Conclusions Iron sucrose incubation is a safe alternative method for ASCs labeling and tracking using MRI following treatment. Clinicians and researchers should be able to visualize the location of ASCs engraftment without secondary surgical investigation involving tissue sampling.
In recent years, a lot of attention has been focused on using adipose‐derived mesenchymal stem cells obtained from infrapatellar fat pad (IPFP‐ASCs) for the articular cartilage regeneration. IPFP‐ASCs constructs were previously characterized and demonstrated chondrogenic differentiation potential to produce hyaline like‐cartilage in vitro. However, little is known about the relationship of its regeneration potential and pain associated with osteochondral defect. This study aimed to investigate the effect of implantation of the 3‐Dimensional (3D) cartilage construct of IPFP‐ASCs on the restoration of an articular hyaline cartilage as well as attenuation of pain associated with the cartilage defect in an osteochondral defect rat model. The chondrogenic differentiation potential of the 3D cartilage construct derived from IPFP‐ASCs was determined prior to implantation and at 4, 8 and 12 weeks post‐implantation by gene expression and immunochemistry analysis. Pain‐related behavior was examined weekly up to 8 weeks post‐implantation by using weight‐bearing test. A significant pain‐associated with osteochondral defect was observed in this model in all groups post‐induction; however, this pain can spontaneously resolve within three weeks post‐implantation regardless of implantation of IPFP‐ASCs constructs. The existences of mature chondrocytes as well as a significant (p<0.05) positively immunostained for type II collagen and aggrecan were identified in the implanted site for up to 12 weeks compared to untreated group, indicating the hyaline cartilage regeneration. Overall, this study reported the successful outcome of osteochondral regeneration with scaffold‐free IPFP‐ASCs constructs in an osteochondral defect rat model. Although the implantation of the cartilage construct could not attenuate pain associated with the cartilage defect, it provides novel and interesting insights into the current hypothesis that 3D construct IPFP‐ASCs may have potential benefits as an alternative approach to repair bone and cartilage defect.
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