Sharun Khan, Pawde Abhijit M, Kumar Rohit, Kalaiselvan E, Kinjavdekar Prakash, Dhama Kuldeep, Pal Amar
Division of Surgery, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India.
Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India.
Vet World. 2021 Feb;14(2):508-514. doi: 10.14202/vetworld.2021.508-514. Epub 2021 Feb 25.
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.
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.
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±0.09×10 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×10 cells/μL.
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.
脂肪组织来源的基质血管成分（SVF）包含由多能脂肪来源干细胞组成的异质细胞群。由于SVF具有卓越的成骨潜力，利用脂肪来源的SVF进行再生治疗在骨组织工程中具有广泛应用。本研究旨在标准化并表征从新西兰白兔获取的脂肪来源SVF，用于骨组织工程及其他潜在应用。
本研究使用了10只骨骼成熟且临床健康的成年新西兰白兔。采用手术切除肩胛间脂肪组织，并用0.1% I型胶原酶溶液进行酶消化来制备SVF。在最后离心步骤后获得的SVF沉淀悬浮于0.5 mL对照溶液中，以获得即用型脂肪来源SVF。基于每克脂肪组织的总SVF细胞计数和细胞产量对新鲜制备的SVF进行表征。使用血细胞计数器对SVF细胞进行计数。
肩胛间脂肪组织库因易于获取，是制备自体脂肪来源SVF的理想选择。通过酶消化制备的肩胛间脂肪来源SVF的平均细胞产量为3.15±0.09×10⁶个细胞/克脂肪组织。新鲜制备的SVF的总细胞计数为3.15±0.09×10⁶个细胞/微升。
使用0.1%胶原酶对脂肪组织进行酶消化，每克脂肪组织的细胞产量比之前在兔子中报道的方法更高。使用脂肪来源的SVF可避免额外的培养期。此外，它还可能降低广泛细胞污染的风险，这使其成为骨组织工程应用中一种安全且经济有效的策略。
