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Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate
Published on: October 31, 2012
A Novel Laboratory-Based Strategy for Single Adipocyte and Adipose-Derived Stem Cells Extraction for Transplantation:
Yu Liu1, Hanxing Zhao1, Jiaying Yu2
1Department of Burn and Plastic Surgery, West China Hospital of Sichuan University, No.37 Guoxue Alley, Wuhou District, Chengdu City, Sichuan Province, People's Republic of China.
Background:
Autologous fat grafting is widely used in plastic surgery. However, its main limitation is the low survival rate of fat grafts after transplantation. Transplantation of single adipocytes in combination with adipose-derived stem cells (ADSCs) could largely preserve the activity of the fat and improve graft survival.
Objective:
To verify the long-term survival rate of single adipocyte graft in vivo and its viable fat morphology for future fat grafting.
Methods:
Healthy adipose tissue was harvested and disassociated using fat dissociation solution, the Single-cell Suspension Preparation System (SSPS) was used to obtain a mixture of single adipocytes, ADSCs and stromal vascular fraction (SVF), and the structure of single adipocytes was verified by cell mask red and DAPI double staining. Nine male Balb/c nude mice were used, and three different graft volumes were established (0.05, 0.1 and 0.2 ml). For each mouse, four sites were selected for transplantation, one for macrofat and the other three for single adipocytes, and different transplant volumes 30, 60 and 90 days after transplantation. In each period, 3 mice were selected to measure the volume of fat graft.
Results:
Double staining with cell mask red and DAPI confirmed that the nucleus was identified intracellularly, which also indicated that the adipocytes in the single-cell suspension were structurally complete. When evaluating the transplantation, the groups with a volume of 0.05 ml and 0.2 ml performed better in the single-cell fat group in all transplantation periods, the group with a volume of 0.1 ml performed better in the single-cell group in the 30- and 60-day transplantation, and the differences were significant (P<0.05).
Conclusion:
In this study, the SSPS was used to obtain a new transplant material containing single adipocytes and ADSCs by enzymatic hydrolysis of adipose tissue and converted into single cells. It effectively improved the survival rate of fat grafting and the long-term effect of transplantation.
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