PMID- 29806371 OWN - NLM STAT- MEDLINE DCOM- 20181005 LR - 20231107 IS - 1002-1892 (Print) IS - 1002-1892 (Linking) VI - 32 IP - 1 DP - 2018 Jan 15 TI - [The role of Schwann cells-like cells derived from human amniotic membrane mesenchymal stem cells transplantation in flap nerves regeneration]. PG - 80-90 LID - 10.7507/1002-1892.201708007 [doi] AB - OBJECTIVE: Inducing human amniotic membrane mesenchymal stem cells (hAMSCs) to Schwann cells-like cells (SCs-like cells) in vitro, and to evaluate the efficacy of transplantation of hAMSCs and SCs-like cells on nerves regeneration of the rat flaps. METHODS: hAMSCs were isolated from placenta via two-step digestion and cultured by using trypsin and collagenase, then identified them by flow cytometry assay and immunofluorescence staining. The 3rd generation of hAMSCs cultured for 6 days were induced to SCs-like cells in vitro; at 19 days after induction, the levels of S-100, p75, and glial fibrillary acidic protein (GFAP) were detected by immunofluorescence staining, Western blot, and real-time fluorescence quantitative PCR (qPCR). The levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were measured by ELISA in the supernatant of the 3rd generation of hAMSCs cultured for 6 days and the hAMSCs induced within 19 days. In addition, 75 female Sprague Dawley rats were taken to establish the rat denervated perforator flap model of the abdominal wall, and were divided into 3 groups ( n=25). The 3rd generation of hAMSCs (1x10 (6) cells) in the proliferation period of culturing for 6 days, the SCs-like cells (1x10 (6) cells), and equal volume PBS were injected subcutaneously in the skin flap of the rat in groups A, B, and C, respectively. At 2, 5, 7, 9, and 14 days after transplantation, 5 rats in each group were killed to harvest the flap frozen sections and observe the positive expression of neurofilament heavy polypeptide antibody (NF-01) by immunofluorescence staining. RESULTS: The cells were identified as hAMSCs by flow cytometry assay and immunofluorescence staining. The results of immunofluorescence staining, Western blot, qPCR showed that the percentage of positive cells, protein expression, and gene relative expression of S-100, p75, and GFAP in SCs-like cells group were significantly higher than those in hAMSCs group ( P<0.05). The results of ELISA demonstrated that the expression of BDNF and NGF was significantly decreased after added induced liquid 1, and the level of BDNF and NGF increased gradually with the induction of liquids 2 and 3, and the concentration of BDNF and NGF was significantly higher than that of hAMSCs group ( P<0.05). Immunofluorescence staining showed that the number of regenerated nerve fibers in group B was higher than that in groups A and C after 5-14 days of transplantation. CONCLUSION: The hAMSCs can be induced into SCs-like cells with the proper chemical factor regulation in vitro, and a large number of promoting nerve growth factor were released during the process of differentiation, and nerve regeneration in flaps being transplanted the SCs-like cells was better than that in flaps being transplanted the hAMSCs, which through a large number of BDNF and NGF were released. FAU - Gong, Feiyu AU - Gong F AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. FAU - Wei, Zairong AU - Wei Z AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China.zairongwei@sina.com. FAU - Jin, Wenhu AU - Jin W AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. FAU - Li, Hai AU - Li H AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. FAU - Deng, Chengliang AU - Deng C AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. FAU - Wu, Bihua AU - Wu B AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. FAU - Nie, Kaiyu AU - Nie K AD - Department of Burn and Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi Guizhou, 563000, P.R.China. LA - chi PT - Journal Article PL - China TA - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi JT - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JID - 9425194 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (NGF protein, human) RN - 7171WSG8A2 (BDNF protein, human) RN - 9061-61-4 (Nerve Growth Factor) SB - IM MH - *Amnion MH - Animals MH - Brain-Derived Neurotrophic Factor MH - Cell Count MH - Cell Differentiation MH - Cells, Cultured MH - Female MH - Humans MH - Mesenchymal Stem Cell Transplantation MH - *Mesenchymal Stem Cells MH - Nerve Growth Factor MH - *Nerve Regeneration MH - Rats MH - Rats, Sprague-Dawley MH - *Schwann Cells MH - Stem Cells/*cytology MH - *Tissue Engineering PMC - PMC8414214 OTO - NOTNLM OT - Human amniotic membrane mesenchymal stem cells OT - Schwann cells-like cells OT - flap OT - induced differentiation OT - nerve regeneration EDAT- 2018/05/29 06:00 MHDA- 2018/10/06 06:00 PMCR- 2018/01/01 CRDT- 2018/05/29 06:00 PHST- 2018/05/29 06:00 [entrez] PHST- 2018/05/29 06:00 [pubmed] PHST- 2018/10/06 06:00 [medline] PHST- 2018/01/01 00:00 [pmc-release] AID - zgxfcjwkzz-32-1-80 [pii] AID - 10.7507/1002-1892.201708007 [doi] PST - ppublish SO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2018 Jan 15;32(1):80-90. doi: 10.7507/1002-1892.201708007.