Standardization and characterization of adipose-derived stromal vascular fraction from New Zealand white rabbits for bone tissue engineering

Abstract

Background and Aim: Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications. Materials and Methods: Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer. Results: Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/μL. Conclusion: The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications.