HUMAN ADIPOSE-DERIVED STEM CELLS COMBINED WITH SMALL INTESNITAL SUBMUCOSA POWDER/CHITOSAN CHLORIDE-β-GLYCEROL PHOSPHATE DISODIUM-HYDROXYETHYL CELLULOSE HYBRID FOR ADIPOSE TISSUE ENGINEERING_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANGShu ^1,2 ,  LUOjingcong ¹ ,  LÜQing ² , DENGXueqin ¹ , XIONGBingjun ^1,2

1. Laboratory of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, Chengdu Sichuan, 610041, P. R. China;
1. Department of Breast Surgery, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;

Corresponding author：

LUOjingcong, Email: ljchx2005@163.com; LÜQing, Email: lqlq1963@163.com

Keywords：

Adipose tissue engineering; Adipose-derived stem cells; Decellular extracellular matrix; Small intestinal submucosa; Nude mice

DOI：

10.7507/1002-1892.20150219

Video：

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Objective To study the feasibility of human adipose-derived stem cells (hADSCs) combined with small intestinal submucosa powder (SISP)/chitosan chloride (CSCl)-β-glycerol phosphate disodium (GP)-hydroxyethyl cellulose (HEC) for adipose tissue engineering. Methods hADSCs were isolated from human breast fat with collagenase type I digestion, and the third passage hADSCs were mixed with SISP/CSCl-GP-HEC at a density of 1×106 cells/mL. Twenty-four healthy female nude mice of 5 weeks old were randomly divided into experimental group (n=12) and control group (n=12), and the mice were subcutaneously injected with 1 mL hADSCs+SISP/CSCl-GP-HEC or SISP/CSCl-GP-HEC respectively at the neck. The degradation rate was evaluated by implant volume measurement at 0, 1, 2, 4, and 8 weeks. Three mice were euthanized at 1, 2, 4, and 8 weeks respectively for general, histological, and immunohistochemical observations. The ability of adipogenesis (Oil O staining), angiopoiesis (CD31), and localized the hADSCs (immunostaining for human Vimentin) were identified. Results The volume of implants of both groups decreased with time, but it was greater in experimental group than the control group, showing significant difference at 8 weeks (t=3.348, P=0.029). The general observation showed that the border of implants was clear with no adhesion at each time point;fat-liked new tissues were observed with capillaries on the surface at 8 weeks in 2 groups. The histological examinations showed that the structure of implants got compact gradually after injection, and SISP gradually degraded with slower degradation speed in experimental group;adipose tissue began to form, and some mature adipose tissue was observed at 8 weeks in the experimental group. The Oil O staining positive area of experimental group was greater than that of the control group at each time point, showing significant difference at 8 weeks (t=3.411, P=0.027). Immunohistochemical staining for Vemintin showed that hADSCs could survive at each time point in the experimental group;angiogenesis was most remarkable at 2 weeks, showing no significant differences in CD31 possitive area between 2 groups (P>0.05), but angiogenesis was more homogeneous in experimental group. Conclusion SISP/CSCl-GP-HEC can use as scaffolds for hADSCs to reconstruct tissue engineered adipose.

Citation： ZHANGShu, LUOjingcong, LÜQing, DENGXueqin, XIONGBingjun. HUMAN ADIPOSE-DERIVED STEM CELLS COMBINED WITH SMALL INTESNITAL SUBMUCOSA POWDER/CHITOSAN CHLORIDE-β-GLYCEROL PHOSPHATE DISODIUM-HYDROXYETHYL CELLULOSE HYBRID FOR ADIPOSE TISSUE ENGINEERING. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(8): 1028-1033. doi: 10.7507/1002-1892.20150219 Copy

1.	Flynn L,Woodhouse KA.Adipose tissue engineering with cells in engineered matrices.Orgnogesis,2008,4(4):228-235.
2.	Kim EH,Heo CY.Current applications of adipose-derived stem cells and their future perspectives.World J Stem Cells,2014,6(1):65-68.
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8.	Zhou Y,Yan Z,Zhang H,et al.Expansion and delivery of adipose-derived mesenchymal stem cells on three microcarriers for soft tissue regeneration.Tissue Eng Part A,2011,17(23-24):2981-2997.
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10.	四川大学华西医院.一种温敏性复合材料及其制备方法和用途:中国,A61L 27/36,2013106019736.2013-11-25.
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14.	Douglas TE,Skwarczynska A,Modrzejewska Z,et al.Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase.Int J Biol Macromol,2013,56:122-132.
15.	刘家宏.胆管癌细胞系QBC939裸鼠神经浸润模型的建立及5'-AMP对裸鼠胆管癌移植瘤作用的实验研究.青岛:青岛大学,2011.
16.	Galateanu B,Dimonie D,Vasile E,et al.Layer-shaped alginate hydrogels enhance the biological performance of human adipose-derived stem cells.BMC Biotechnol,2012,12:35.
17.	Choi YS,Cha SM,Lee YY,et al.Adipogenic differentiation of adipose tissue derived adult stem cells in nude mouse.Biochem Biophys Res Commun,2006,345(2):631-637.
18.	Zimmerlin L,Donnenberg AD,Rubin JP,et al.Regenerative therapy and cancer:in vitro and in vivo studies of the interaction between adipose-derived stem cells and breast cancer cells from clinical isolates.Tissue Eng Part A,2011,17(1-2):93-106.
19.	Dolderer JH,Medved F,Haas RM,et al.Angiogenesis and vascularisation in adipose tissue engineering.Handchir Mikrochir Plast Chir,2013,45(2):99-107.
20.	Wang L,Johnson JA,Zhang Q,et al.Combining decellularized human adipose tissue extracellular matrix and adipose-derived stem cells for adipose tissue engineering.Acta Biomater,2013,9(11):8921-8931.
21.	Fumimoto Y,Matsuyama A,Komoda H,et al.Creation of a rich subcutaneous vascular network with implanted adipose tissue-derived stromal cells and adipose tissue enhances subcutaneous grafting of islets in diabetic mice.Tissue Eng Part C Methods,2009,15(3):437-444.
22.	Kim WS,Park BS,Sung JH,et al.Wound healing effect of adipose-derived stem cells:a critical role of secretory factors on human dermal fibroblasts.J Dermatol Sci,2007,48(1):15-24.
23.	Li CJ,Madhu V,Balian G,et al.Cross-talk between VEGF and BMP-6 pathways accelerates osteogenic differentiation of human adipose-derived stem cells.J Cell Physiol,2015.[Epub ahead of print].
24.	Vallée M,Côté JF,Fradette J.Adipose-tissue engineering:taking advantage of the properties of human adipose-derived stem/stromal cells.Pathol Biol (Paris),2009,57(4):309-317.
25.	Hausman GJ,Richardson RL.Adipose tissue angiogenesis.J Anim Sci,2004,82(3):925-934.

1. Flynn L,Woodhouse KA.Adipose tissue engineering with cells in engineered matrices.Orgnogesis,2008,4(4):228-235.
2. Kim EH,Heo CY.Current applications of adipose-derived stem cells and their future perspectives.World J Stem Cells,2014,6(1):65-68.
3. Huang SJ,Fu RH,Shyu WC,et al.Adipose-derived stem cells:isolation,characterization,and differentiation potential.Cell Transplant,2013,22(4):701-709.
4. Deng M,Gu Y,Liu Z,et al.Endothelial differentiation of human adipose-derived stem cells on polyglycolic acid/polylactic acid mesh.Stem Cells Int,2015,2015:350718.
5. Gimble JM,Katz AJ,Bunnell BA.Adipose-derived stem cells for regenerative medicine.Circ Res,2007,100(9):1249-1260.
6. Porzionato A,Sfriso MM,Pontini A,et al.Decellularized human skeletal muscle as biologic scaffold for reconstructive surgery.Int J Mol Sci,2015,16(7):14808-14831.
7. Rana D,Zreiqat H,Benkirane-Jessel N,et al.Development of decellularized scaffolds for stem cell-driven tissue engineering.J Tissue Eng Regen Med,2015.[Epub ahead of print].
8. Zhou Y,Yan Z,Zhang H,et al.Expansion and delivery of adipose-derived mesenchymal stem cells on three microcarriers for soft tissue regeneration.Tissue Eng Part A,2011,17(23-24):2981-2997.
9. 邓雪琴.猪小肠黏膜下层微粉/氯化壳聚糖杂化温敏材料初步研究.成都:四川大学,2013.
10. 四川大学华西医院.一种温敏性复合材料及其制备方法和用途:中国,A61L 27/36,2013106019736.2013-11-25.
11. Flynn L,Semple JL,Woodhouse KA.Decellularized placental matrices for adipose tissue engineering.J Biomed Mater Res A,2006,79(2):359-369.
12. 田春祥,范雪娇,陈晓禾,等.骨骼肌无细胞基质的制备及其生物相容性研究.中国修复重建外科杂志,2012,26(6):749-754.
13. Hillel AT,Nahas Z,Unterman S,et al.Validation of a small animal model for soft tissue filler characterization.Dermatol Surg,2012,38(3):471-478.
14. Douglas TE,Skwarczynska A,Modrzejewska Z,et al.Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase.Int J Biol Macromol,2013,56:122-132.
15. 刘家宏.胆管癌细胞系QBC939裸鼠神经浸润模型的建立及5'-AMP对裸鼠胆管癌移植瘤作用的实验研究.青岛:青岛大学,2011.
16. Galateanu B,Dimonie D,Vasile E,et al.Layer-shaped alginate hydrogels enhance the biological performance of human adipose-derived stem cells.BMC Biotechnol,2012,12:35.
17. Choi YS,Cha SM,Lee YY,et al.Adipogenic differentiation of adipose tissue derived adult stem cells in nude mouse.Biochem Biophys Res Commun,2006,345(2):631-637.
18. Zimmerlin L,Donnenberg AD,Rubin JP,et al.Regenerative therapy and cancer:in vitro and in vivo studies of the interaction between adipose-derived stem cells and breast cancer cells from clinical isolates.Tissue Eng Part A,2011,17(1-2):93-106.
19. Dolderer JH,Medved F,Haas RM,et al.Angiogenesis and vascularisation in adipose tissue engineering.Handchir Mikrochir Plast Chir,2013,45(2):99-107.
20. Wang L,Johnson JA,Zhang Q,et al.Combining decellularized human adipose tissue extracellular matrix and adipose-derived stem cells for adipose tissue engineering.Acta Biomater,2013,9(11):8921-8931.
21. Fumimoto Y,Matsuyama A,Komoda H,et al.Creation of a rich subcutaneous vascular network with implanted adipose tissue-derived stromal cells and adipose tissue enhances subcutaneous grafting of islets in diabetic mice.Tissue Eng Part C Methods,2009,15(3):437-444.
22. Kim WS,Park BS,Sung JH,et al.Wound healing effect of adipose-derived stem cells:a critical role of secretory factors on human dermal fibroblasts.J Dermatol Sci,2007,48(1):15-24.
23. Li CJ,Madhu V,Balian G,et al.Cross-talk between VEGF and BMP-6 pathways accelerates osteogenic differentiation of human adipose-derived stem cells.J Cell Physiol,2015.[Epub ahead of print].
24. Vallée M,Côté JF,Fradette J.Adipose-tissue engineering:taking advantage of the properties of human adipose-derived stem/stromal cells.Pathol Biol (Paris),2009,57(4):309-317.
25. Hausman GJ,Richardson RL.Adipose tissue angiogenesis.J Anim Sci,2004,82(3):925-934.

Chinese Journal of Reparative and Reconstructive Surgery

HUMAN ADIPOSE-DERIVED STEM CELLS COMBINED WITH SMALL INTESNITAL SUBMUCOSA POWDER/CHITOSAN CHLORIDE-β-GLYCEROL PHOSPHATE DISODIUM-HYDROXYETHYL CELLULOSE HYBRID FOR ADIPOSE TISSUE ENGINEERING

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