EFFECT OF HUMAN PLACENTAL DECIDUA BASALIS DERIVED MESENCHYMAL STEM CELLS IN REPAIR OF NUDE MICE SKIN WOUND_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

SUNNing ¹ , HUANGYizhou ¹ , ZHANGYi ² , PIJinkui ¹ , LawJaznie ³ ,  DENGLi ¹ ,  WUChengguang ¹

1. Laboratory of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;
2. Core Facility, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;
3. Department of Burn and Plastic Surgery, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;

Corresponding author：

DENGLi, Email: dengli2000@gmail.com; WUChengguang, Email: chengguang1989@foxmail.com

Keywords：

Mesenchymal stem cells; Placenta; Skin defect; Repair and reconstruction; Nude mice

DOI：

10.7507/1002-1892.20160125

Video：

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Objective To investigate the effectiveness of human placental decidua basalis derived mesenchymal stem cells (PDB-MSCs) in repairing full-thickness skin defect of nude mice. Methods Human placenta samples were obtained from healthy donor mothers with written informed consent. PDB-MSCs were isolated through enzymic digestion and density gradient centrifugation; the 4th passage cells were identified by cellular morphology, cell adipogenic and osteogenic differentiation, and phenotype evaluation. Forty-two 4-5-week-old BALB/c female nude mice were randomly divided into experimental group (n=21) and control group (n=21). The 4th passage PDB-MSCs solution (200 μL, 5×10⁶/mL) was injected into the mice of experimental group via caudal vein; the mice of control group were given equal volume of PBS. The full-thickness skin defect model of 1.5 cm×1.5 cm in size was made after 3 days. The wound healing was observed generally at 1, 2, 4, 7, 14, 18, 21, 25, and 30 days after operation, and the wound healing rate was calculated after wound decrustation. HE staining was used to observe the wound repair at 1, 7, 14, 21, and 31 days; immunofluorescent staining was used for cellular localization at 7, 14, and 31 days after operation. Results Cells isolated from human placenta were MSCs which had multipotential differentiation ability and expressed MSCs phenotype. Animals survived to the end of the experiment. The general observation showed that the experimental group had a faster skin repairing speed than the control group; the time for decrustation was 12-14 days in experimental group and was 14-17 days after operation in the control group. The wound healing rate of experimental group was significantly higher than that of control group at 14, 18, and 21 days (t=4.001, P=0.016; t=3.380, P=0.028; t=3.888, P=0.018), but no significance was found at 25 and 30 days (t=1.565, P=0.193; t=1.000, P=0.423). HE staining showed lower inflammatory reaction, and better regeneration of the whole skin and glands with time in the experimental group. The immunofluorescent staining was positive in skin defect area of experimental group at different time points which displayed that human PDB-MSCs existed. Conclusion Through enzymic digestion and density gradient centrifugation, PDB-MSCs can be obtained. Pre-stored PDB-MSCs can mobilize to the defect area and participate in repair of nude mice skin.

Citation： SUNNing, HUANGYizhou, ZHANGYi, PIJinkui, LawJaznie, DENGLi, WUChengguang. EFFECT OF HUMAN PLACENTAL DECIDUA BASALIS DERIVED MESENCHYMAL STEM CELLS IN REPAIR OF NUDE MICE SKIN WOUND. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(5): 619-625. doi: 10.7507/1002-1892.20160125 Copy

1.	Li H, Fu X, Ouyang Y, et al. Adult bone-marrow-derived mesenchymal stem cells contribute to wound healing of skin appendages. Cell Tissue Res, 2006, 326(3):725-736.
2.	Fu X, Fang L, Li X, et al. Enhanced wound-healing quality with bone marrow mesenchymal stem cells autografting after skin injury. Wound Repair Regen, 2006, 14(3):325-335.
3.	Wu CG, Zhang JC, Xie CQ, et al. In vivo tracking of human placenta derived mesenchymal stem cells in nude mice via ¹⁴C-TdR labeling. BMC Biotechnology, 2015, 15(55):DOI:10.1186/s12896-015-0174-4.
4.	Li ZH, Liao W, Cui XL, et al. Intravenous transplanlation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci, 2011, 8(1):74-83.
5.	Oh SH, Muzzonigro TM, Bae SH, et al. Adult bone marrow-derived cells trans-differentiating in to insulin-producing cells for the treatment of type I diabetes. Lab Invest, 2004, 84(5):607-617.
6.	Ende N, Chen R, Reddi AS. Effect of human umbilical cord blood cells on glycemia and insulitis in type 1 diabetic mice. Biochem Biophys Res Commun, 2004, 325(3):665-669.
7.	Lim JH, Byeon YE, Ryu HH, et al. Transplantation of canine umbilical cord blood-derived mesenchymal stem cells in experimentally induced spinal cord injured dogs. J Vet Sci, 2007, 8(3):275-282.
8.	Coutts M, Keirstead HS. Stem cells for the treatment of spinal cord injury. Exp Neurol, 2008, 209(2):368-377.
9.	Pereira WC, Khushnooma I, Madkaikar M, et al. Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation. J Tissue Eng Regen Med, 2008, 2(7):394-399.
10.	Ichim TE, Alexandrescu DT, Solano F, et al. Mesenchymal stem cells as anti-infl ammatories:implications for treatment of Duchennemuscular dystrophy. Cell Immunol, 2010, 260(2):75-82.
11.	Magatti M, De Munari S, Vertua E, et al. Human amnion mesenchyme harbors cells with allogeneic T-cell suppression and stimulation capabilities. Stem Cells, 2008, 26(1):182-192.
12.	Parolini O, Caruso M. Review:Preclinical studies on placentaderived cells and amniotic membrane:an update. Placenta, 2011, 32 Suppl 2:S186-195.
13.	Miao Z, Jin J, Chen L, et al. Isolation of mesenchymal stem cells from human placenta:comparison with human bone marrow mesenchymal stem cells. Cell Biol Int, 2006, 30(9):681-687.
14.	Chien CC, Yen BL, Lee FK, et al. In vitro differentiation of human placenta-derived multipotent cells into hepatocyte-like cells. Stem Cells, 2006, 24(7):1759-1768.
15.	Chang CM, Kao CL, Chang YL, et al. Placenta-derived multipotent stem cells induced to differentiate into insulin-positive cells. Biochem Biophys Res Commun, 2007, 357(2):414-420.
16.	Krause DS, Theise ND, Collector MI, et al. Multi-organ, multilineage engraftment by a single bone marrow-derived stem cell. Cell, 2001, 105(3):369-377.
17.	Kwon DS, Gao X, Liu YB, et al. Treatment with bone marrowderived stromal cells accelerates wound healing in diabetic rats. Int Wound J, 2008, 5(3):453-463.
18.	Kataoka K, Medina RJ, Kageyama T, et al. Participation of adult mouse bone marrow cells in reconstitution of skin. Am J Pathol, 2003, 163(4):1227-1231.
19.	Satoh H, Kishi K, Tanaka T, et al. Transplanted mesenchymal stem cells are effective for skin regeneration in acute cutaneous wounds. Cell Transplant, 2004, 13(4):405-412.
20.	Zou ZM, Yang Y, Hao L, et al. More insight into mesenchymal stem cells and their effects inside the body. Expert Opin Biol Ther, 2010, 10(2):215-230.
21.	Liang X, Ding Y, Zhang Y, et al. Paracrine mechanisms of mesenchymal stem cell-based therapy:current status and perspectives. Cell Transplantation, 2014, 23(9):1045-1059.
22.	Ermolov AS, Smirnov SV, Khvatov VB, et al. The use of bioactive wound dressing, stimulating epithelial regeneration of IIIa-degree burn wounds. Bull Exp Biol Med, 2008, 146(1):153-157.
23.	Pallua N, Pulsfort AK, Suschek C, et al. Content of the growth factors bFGF, IGF-1, VEGF, and PDGF-BB in freshly harvested lipoaspirate after centrifugation and incubation. Plast Reconstr Surg, 2009, 123(3):826-833.
24.	Ching YH, Sutton TL, Pierpont YN, et al. The use of growth factors and other humoral agents to accelerate and enhance burn wound healing. Eplasty, 2011, 11:e41.

1. Li H, Fu X, Ouyang Y, et al. Adult bone-marrow-derived mesenchymal stem cells contribute to wound healing of skin appendages. Cell Tissue Res, 2006, 326(3):725-736.
2. Fu X, Fang L, Li X, et al. Enhanced wound-healing quality with bone marrow mesenchymal stem cells autografting after skin injury. Wound Repair Regen, 2006, 14(3):325-335.
3. Wu CG, Zhang JC, Xie CQ, et al. In vivo tracking of human placenta derived mesenchymal stem cells in nude mice via ¹⁴C-TdR labeling. BMC Biotechnology, 2015, 15(55):DOI:10.1186/s12896-015-0174-4.
4. Li ZH, Liao W, Cui XL, et al. Intravenous transplanlation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci, 2011, 8(1):74-83.
5. Oh SH, Muzzonigro TM, Bae SH, et al. Adult bone marrow-derived cells trans-differentiating in to insulin-producing cells for the treatment of type I diabetes. Lab Invest, 2004, 84(5):607-617.
6. Ende N, Chen R, Reddi AS. Effect of human umbilical cord blood cells on glycemia and insulitis in type 1 diabetic mice. Biochem Biophys Res Commun, 2004, 325(3):665-669.
7. Lim JH, Byeon YE, Ryu HH, et al. Transplantation of canine umbilical cord blood-derived mesenchymal stem cells in experimentally induced spinal cord injured dogs. J Vet Sci, 2007, 8(3):275-282.
8. Coutts M, Keirstead HS. Stem cells for the treatment of spinal cord injury. Exp Neurol, 2008, 209(2):368-377.
9. Pereira WC, Khushnooma I, Madkaikar M, et al. Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation. J Tissue Eng Regen Med, 2008, 2(7):394-399.
10. Ichim TE, Alexandrescu DT, Solano F, et al. Mesenchymal stem cells as anti-infl ammatories:implications for treatment of Duchennemuscular dystrophy. Cell Immunol, 2010, 260(2):75-82.
11. Magatti M, De Munari S, Vertua E, et al. Human amnion mesenchyme harbors cells with allogeneic T-cell suppression and stimulation capabilities. Stem Cells, 2008, 26(1):182-192.
12. Parolini O, Caruso M. Review:Preclinical studies on placentaderived cells and amniotic membrane:an update. Placenta, 2011, 32 Suppl 2:S186-195.
13. Miao Z, Jin J, Chen L, et al. Isolation of mesenchymal stem cells from human placenta:comparison with human bone marrow mesenchymal stem cells. Cell Biol Int, 2006, 30(9):681-687.
14. Chien CC, Yen BL, Lee FK, et al. In vitro differentiation of human placenta-derived multipotent cells into hepatocyte-like cells. Stem Cells, 2006, 24(7):1759-1768.
15. Chang CM, Kao CL, Chang YL, et al. Placenta-derived multipotent stem cells induced to differentiate into insulin-positive cells. Biochem Biophys Res Commun, 2007, 357(2):414-420.
16. Krause DS, Theise ND, Collector MI, et al. Multi-organ, multilineage engraftment by a single bone marrow-derived stem cell. Cell, 2001, 105(3):369-377.
17. Kwon DS, Gao X, Liu YB, et al. Treatment with bone marrowderived stromal cells accelerates wound healing in diabetic rats. Int Wound J, 2008, 5(3):453-463.
18. Kataoka K, Medina RJ, Kageyama T, et al. Participation of adult mouse bone marrow cells in reconstitution of skin. Am J Pathol, 2003, 163(4):1227-1231.
19. Satoh H, Kishi K, Tanaka T, et al. Transplanted mesenchymal stem cells are effective for skin regeneration in acute cutaneous wounds. Cell Transplant, 2004, 13(4):405-412.
20. Zou ZM, Yang Y, Hao L, et al. More insight into mesenchymal stem cells and their effects inside the body. Expert Opin Biol Ther, 2010, 10(2):215-230.
21. Liang X, Ding Y, Zhang Y, et al. Paracrine mechanisms of mesenchymal stem cell-based therapy:current status and perspectives. Cell Transplantation, 2014, 23(9):1045-1059.
22. Ermolov AS, Smirnov SV, Khvatov VB, et al. The use of bioactive wound dressing, stimulating epithelial regeneration of IIIa-degree burn wounds. Bull Exp Biol Med, 2008, 146(1):153-157.
23. Pallua N, Pulsfort AK, Suschek C, et al. Content of the growth factors bFGF, IGF-1, VEGF, and PDGF-BB in freshly harvested lipoaspirate after centrifugation and incubation. Plast Reconstr Surg, 2009, 123(3):826-833.
24. Ching YH, Sutton TL, Pierpont YN, et al. The use of growth factors and other humoral agents to accelerate and enhance burn wound healing. Eplasty, 2011, 11:e41.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF HUMAN PLACENTAL DECIDUA BASALIS DERIVED MESENCHYMAL STEM CELLS IN REPAIR OF NUDE MICE SKIN WOUND

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