Mesenchymal progenitor cells derived from traumatized human muscle

Mesenchymal stem cells (MSCs) derived from adult tissues are an important candidate cell type for cell‐based tissue engineering and regenerative medicine. Currently, clinical applications for MSCs require additional surgical procedures to harvest the autologous MSCs (i.e. from bone marrow) or commercial allogeneic alternatives. We have recently identified a population of mesenchymal progenitor cells (MPCs) in traumatized muscle tissue that has been surgically debrided from traumatic orthopaedic extremity wounds. The purpose of this study was to evaluate whether MPCs derived from traumatized muscle may provide a clinical alternative to bone‐marrow MSCs, by comparing their morphology, proliferation capacity, cell surface epitope profile and differentiation capacity. After digesting the muscle tissue with collagenase, the MPCs were enriched by a direct plating technique. The morphology and proliferation rate of the muscle‐derived MPCs was similar to bone‐marrow derived MSCs. Both populations expressed cell surface markers characteristic for MSCs (CD 73, CD 90 and CD105), and did not express markers typically absent on MSCs (CD14, CD34 and CD45). After 21 days in specific differentiation media, the histological staining and gene expression of the MPCs and MSCs was characteristic for differentiation into osteoblasts, chondrocytes and adipocytes, but not into myoblasts. Our findings demonstrate that traumatized muscle‐derived MPCs exhibit a similar phenotype and resemble MSCs derived from the bone marrow. MPCs harvested from traumatized muscle tissue may be considered for applications in tissue engineering and regenerative medicine following orthopaedic trauma requiring circumferential debridement. Copyright © 2009 John Wiley & Sons, Ltd.     

骨髓间充质干细胞表达神经营养因子及对神经干细胞的保护作用

目的 研究大鼠骨髓间充质干细胞（BMSC）表达脑源性神经营养因子（BDNF）和神经生长因子（NGF）以及BMSC条件培养液对神经干细胞的保护作用。方法 取大鼠骨髓培养BMSC，用CD44、CD45和CD71免疫细胞化学染色进行鉴定。提取BMSCmRNA，RT-PCR检测BDNF和NGF表达，ELISA检测BMSC培养液中BDNF和NGF的含量。原代取材培养新生大鼠皮质神经干细胞，nestin染色鉴定。用不同比例的BMSC条件培养液培养神经干细胞24h后，热休克诱导凋亡，流式细胞仪检测凋亡细胞比率。结果 BMSC贴壁生长，免疫细胞化学染色CD44和CD71阳性表达．CD45阴性表达。BMSC表达BDNF和NGFmRNA，BMSC培养液中含有BDNF和NGF；随培养时间的延长，培养液中BDNF和NGF的浓度增加。BMSC条件培养液可以减少热休克诱导的神经干细胞凋亡比率．并且BMSC条件培养液浓度高者有更好的保护作用。结论 BMSC表达神经营养因子BDNF和NGF，对神绎千细朐凋亡有保护作用．     

Intraperitoneal administration of adipose tissue‐derived stem cells for the rescue of retinal degeneration in a mouse model via indigenous CNTF up‐regulation by IL‐6

As the world's population begins to age, retinal degeneration is an increasing problem, and various treatment modalities are being developed. However, there have been no therapies for degenerative retinal conditions that are not characterized by neovascularization. We investigated whether transplantation of mouse adipose tissue‐derived stem cells (mADSC) into the intraperitoneal space has a rescue effect on NaIO₃‐induced retinal degeneration in mice. In this study, mADSC transplantation recovered visual function and preserved the retinal outer layer structure compared to the control group without any integration of mADSC into the retina. Moreover, endogenous ciliary neurotrophic factor (CNTF) was elevated in the retinas of mADSC‐treated mice. We found that lipopolysaccharide (LPS) or LPS‐stimulated monocyte supernatant induced the secretion of granulocyte colony stimulating factor (GCSF), CD54, CXCL10, interleukin‐6 (IL‐6), and CCL5 from the mADSC by cytokine array. Network inference was conducted to investigate signaling networks related to CNTF regulation. Based on bioinformatics data, the expression of IL‐6 was related to the expression of CNTF. Additionally, intravitreal injection of IL‐6 in rats produced up‐regulation of endogenous CNTF in the retina. mADSC had a rescue effect on retinal degeneration through the up‐regulation of endogenous CNTF by IL‐6. Thus, transplantation of mADSC could be a potential treatment option for retinal degeneration.     

脑源性神经营养因子对面神经损伤的修复作用

目的探讨局部应用脑源性神经营养因子(BDNF)对面神经损伤的修复作用。方法将成年新西兰兔面神经上颊支切除5mm,在断端间置入硅胶再生室,随机实验组室内注入BDNF,对照组用生理盐水。术后4周和8周进行电生理学、组织病理学观察和形态学定量分析。结果术后4周,两组电刺激面神经很少能引发面肌兴奋,有髓轴索计数,轴索直径和面积t检验,两组差异有显著性意义(P<0.05)。术后8周面神经复合肌动作电位(CMAP)BDNF组运动神经传导速度(MCV)明显短于对照组,有髓轴索计数,轴索直径和面积BDNF组大于对照组(P<0.05)。结论局部应用BDNF可促进面神经损伤的修复。     

Vascular endothelial growth factor‐transfected adipose‐derived stromal cells enhance bone regeneration and neovascularization from bone marrow stromal cells

Vascular endothelial growth factor (VEGF)‐transfected adipose‐derived stromal cells (ADSCs^VEGF) were devised to promote bone regeneration and neovascularization of bone marrow stromal cells (BMSCs). ADSCs^VEGF were added to BMSCs and cocultured in variable proportions. ADSCs^VEGF alone or ADSCs^VEGF with BMSCs (BMSCs:ADSCs^VEGF ratio of 1:0.025–0.5) induced significantly greater tube formation in human umbilical vein endothelial cells than untransfected ADSCs. The cocultures of BMSCs and ADSCs^VEGF at ratios of 1: 0.025–0.1 showed significantly greater osteogenic differentiation and mineralization than BMSCs alone in vitro. Osteogenic markers COL1A1, OCN and BSP were most effectively induced at the BMSC: ADSC^VEGF ratio of 1:0.05. Of angiogenesis‐related genes, upregulation of cathepsin Z and downregulation of early growth response 1 were observed while two osteogenesis‐related genes, osteoactivin and tetranectin, were upregulated in BMSCs/ADSCs^VEGF compared to BMSCs/ADSCs. When critical size calvarial defects in rats were implanted with mixture of BMSCs and ADSCs^VEGF along with hydroxyapatite/β‐tricalcium phosphate granules, BMSCs and ADSCs^VEGF at the ratio of 1:0.05 showed better bone regeneration that BMSCs alone. The cotransplantation of ADSCs^VEGF with BMSCs significantly increased neovascularization on the regenerated bone of the repaired defect than BMSCs alone. In conclusion, ADSCs^VEGF added in small proportion to BMSCs effectively promote bone regeneration and neovascularization. Copyright © 2017 John Wiley & Sons, Ltd.     

The role of brain‐derived neurotrophic factor in different animal models of neuropathic pain

Even in present day pain therapy, neuropathic pain remains a challenge for clinicians to treat and a challenge for researchers to investigate. Different animal models have been developed to mimic neuropathic pain. Neurotrophins such as nerve growth factor, brain‐derived neurotrophic factor and neurotrophin 3 have been studied extensively in these models, yet few review articles concerning brain‐derived neurotrophic factor have been published. This article reassesses the literature concerning brain‐derived neurotrophic factor expression in the sciatic nerve chronic constriction injury model, the sciatic nerve transection model, the spinal nerve ligation model and the spinal nerve transection model and discusses differences in regulation of brain‐derived neurotrophic factor between these models and their causality with neuropathic pain.     

Cultured cell‐derived extracellular matrices to enhance the osteogenic differentiation and angiogenic properties of human mesenchymal stem/stromal cells

Cell‐derived extracellular matrix (ECM) consists of a complex assembly of fibrillary proteins, matrix macromolecules, and associated growth factors that mimic the composition and organization of native ECM micro‐environment. Therefore, cultured cell‐derived ECM has been used as a scaffold for tissue engineering settings to create a biomimetic micro‐environment, providing physical, chemical, and mechanical cues to cells, and support cell adhesion, proliferation, migration, and differentiation. Here, we present a new strategy to produce different combinations of decellularized cultured cell‐derived ECM (dECM) obtained from different cultured cell types, namely, mesenchymal stem/stromal cells (MSCs) and human umbilical vein endothelial cells (HUVECs), as well as the coculture of MSC:HUVEC and investigate the effects of its various compositions on cell metabolic activity, osteogenic differentiation, and angiogenic properties of human bone marrow (BM)‐derived MSCs, vital features for adult bone tissue regeneration and repair. Our findings demonstrate that dECM presented higher cell metabolic activity compared with tissue culture polystyrene. More importantly, we show that MSC:HUVEC ECM enhanced the osteogenic and angiogenic potential of BM MSCs, as assessed by in vitro assays. Interestingly, MSC:HUVEC (1:3) ECM demonstrated the best angiogenic response of MSCs in the conditions tested. To the best of our knowledge, this is the first study that demonstrates that dECM derived from a coculture of MSC:HUVEC impacts the osteogenic and angiogenic capabilities of BM MSCs, suggesting the potential use of MSC:HUVEC ECM as a therapeutic product to improve clinical outcomes in bone regeneration.     

Tissue‐specific composite cell aggregates drive periodontium tissue regeneration by reconstructing a regenerative microenvironment

Periodontitis is the most common cause of periodontium destruction. Regeneration of damaged tissue is the expected treatment goal. However, the regeneration of a functional periodontal ligament (PDL) insertion remains a difficulty, due to complicated factors. Recently, periodontal ligament stem cells (PDLSCs) and bone marrow‐derived mesenchymal stem cells (BMMSCs) have been shown to participate in PDL regeneration, both pathologically and physiologically. Besides, interactions affect the biofunctions of different derived cells during the regenerative process. Therefore, the purpose of this study was to discuss the different derived composite cell aggregate (CA) systems of PDLSCs and BMMSCs (iliac‐derived or jaw‐derived) for periodontium regeneration under regenerative microenvironment reconstruction. Our results showed although all three mono‐MSC CAs were compacted and the cells arranged regularly in them, jaw‐derived BMMSC (JBMMSC) CAs secreted more extracellular matrix than the others. Furthermore, PDLSC/JBMMSC compound CAs highly expressed ALP, Col‐I, fibronectin, integrin‐β1 and periostin, suggesting that their biofunction is more appropriate for periodontal structure regeneration. Inspiringly, PDLSC/JBMMSC compound CAs regenerated more functional PDL‐like tissue insertions in both nude mice ectopic and minipig orthotopic transplantation. The results indicated that the different derived CAs of PDLSCs/JBMMSCs provided an appropriate regenerative microenvironment facilitating a more stable and regular regeneration of functional periodontium tissue. This method may provide a possible strategy to solve periodontium defects in periodontitis and powerful experimental evidence for clinical applications in the future. Copyright © 2015 John Wiley & Sons, Ltd.     

Traumatized muscle‐derived multipotent progenitor cells recruit endothelial cells through vascular endothelial growth factor‐A action

Traumatized muscle, such as that debrided from blast injury sites, is considered a promising and convenient tissue source for multipotent progenitor cells (MPCs), a population of adult mesenchymal stem cell (MSC)‐like cells. The present study aimed to assess the regenerative therapeutic potential of human traumatized muscle‐derived MPCs, e.g., for injury repair in the blast‐traumatized extremity, by comparing their pro‐angiogenic potential in vitro and capillary recruitment activity in vivo to those of MSCs isolated from human bone marrow, a widely‐used tissue source. MPCs were tested for their direct and indirect effects on human microvascular endothelial cells (ECs) in vitro. The findings reported here showed that MPC‐conditioned culture medium (MPC‐CM), like MSC‐CM, promoted EC‐cord network branching. Silent (si)RNA‐mediated silencing of vascular endothelial growth factor‐A (VEGF‐A) expression in MPCs attenuated this effect. In a chick embryonic chorioallantoic membrane in vivo angiogenesis assay, MPCs encapsulated in photocrosslinked gelatin scaffold recruited blood vessels more efficiently than either MSCs or human foreskin fibroblasts. Together, these findings support the potential application of traumatized muscle‐derived MPCs in cell‐based regenerative medicine therapies as a result of their influence on EC organization. Copyright © 2017 John Wiley & Sons, Ltd.     

A promising therapeutic option for diabetic bladder dysfunction: Adipose tissue‐derived stem cells pretreated by defocused low‐energy shock wave

Adipose tissue‐derived stem cells (ADSCs) have shown effectiveness in treating diabetic bladder dysfunction (DBD). In the present study, ADSCs pretreated by defocused low‐energy shock wave (DLSW) were first used to achieve better therapeutic effect. ADSCs were treated by DLSW prior to each passage. Secretions of vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) were tested. Proliferation ability was examined by staining 5‐ethynyl‐2‐deoxyuridine (EdU) and assessing expressions of proliferating cell nuclear antigen (PCNA) and Ki67. DBD rat model was created and subgrouped via therapeutic options of phosphate‐buffered saline, ADSCs, pretreated ADSCs, and ADSCs lysate. Afterward, voiding functions were evaluated, and tissues were examined by histology. Neonatal rats received intraperitoneal injection of EdU. All rats were subgrouped and treated as narrated above. Bladder tissues were stained with EdU, Stro‐1, and CD34. Results showed that shocked ADSCs were activated by secreting more VEGF and NGF, by higher EdU‐retaining cells ratios, and by higher expressions of PCNA and Ki67 compared with unshocked ADSCs. Shocked ADSCs had the most effective efficacy in treating DBD by secreting the most VEGF and NGF to accelerate regenerations of revascularization and innervation. Migrations of EdU⁺Stro‐1⁺CD34^? endogenous stem cells to bladders were enhanced by injecting ADSCs. In conclusion, ADSCs pretreated by DLSW had potent therapeutic effect in treating DBD by secreting VEGF and NGF. Recruitment of endogenous stem cells was considered as an important mechanism in this regenerative process.     

Human adipose tissue‐derived tenomodulin positive subpopulation of stem cells: A promising source of tendon progenitor cells

Cell‐based therapies are of particular interest for tendon and ligament regeneration given the low regenerative potential of these tissues. Adipose tissue is an abundant source of stem cells, which may be employed for the healing of tendon lesions. However, human adult multipotent adipose‐derived stem cells (hASCs) isolated from the stromal vascular fraction of adipose tissue originate highly heterogeneous cell populations that hinder their use in specific tissue‐oriented applications. In this study, distinct subpopulations of hASCs were immunomagnetic separated and their tenogenic differentiation capacity evaluated in the presence of several growth factors (GFs), namely endothelial GF, basic‐fibroblast GF, transforming GF‐β1 and platelet‐derived GF‐BB, which are well‐known regulators of tendon development, growth and healing. Among the screened hASCs subpopulations, tenomodulin‐positive cells were shown to be more promising for tenogenic applications and therefore this subpopulation was further studied, assessing tendon‐related markers (scleraxis, tenomodulin, tenascin C and decorin) both at gene and protein level. Additionally, the ability for depositing collagen type I and III forming extracellular matrix structures were weekly assessed up to 28 days. The results obtained indicated that tenomodulin‐positive cells exhibit phenotypical features of tendon progenitor cells and can be biochemically induced towards tenogenic lineage, demonstrating that this subset of hASCs can provide a reliable source of progenitor cells for therapies targeting tendon regeneration.     

自体骨髓间质干细胞移植治疗缺血性脑损伤的实验研究

目的:研究自体骨髓间质干细胞移植对大鼠缺血性脑损伤的治疗作用及机制。方法:采用线拴法制作成年雄性SD大鼠脑缺血再灌注模型40只,随机分成磷酸盐缓冲液对照组和骨髓间质干细胞移植治疗组各20只,分别在1周后经颈动脉将标记5-溴脱氧尿嘧啶的2×106骨髓间质干细胞悬液和等体积磷酸盐缓冲液植入脑缺血大鼠体内。术后每天进行神经功能缺损评分,再灌注后1,2,3,4周分别取受损伤脑组织,检测不同时间点体感诱发电位与组织病理学,并采用免疫组化方法检测脑源性神经营养因子的表达。结果:治疗组在移植后各时间点神经功能缺损评分均明显低于对照组(P<0.05);治疗组神经细胞变性坏死数量减少,水肿明显减轻,体感诱发电位在各时间点的恢复较对照组差异均有统计学意义(P<0.05);治疗组脑源性神经营养因子表达水平明显高于对照组(P<0.05)。结论:自体骨髓间质干细胞移植对大鼠缺血性脑损伤有治疗作用,其机制可能与分泌脑源性神经营养因子等神经保护性因子有关。     

超短波对大鼠周围神经缺损修复术后神经再生的影响*

目的：通过检测小剂量超短波对脱细胞同种异体神经移植物修复大鼠坐骨神经缺损后再生神经传导速度、患侧胫前肌湿重比率、患侧L4脊髓内脑源性神经营养因子（BDNF）和降钙素基因相关肽（CGRP）的表达,来观察小剂量超短波对周围神经缺损修复术后神经再生的影响。方法：将36只Wistar大鼠随机分成3组：正常组（n=4）,对照组（n=16）及实验组（n=16）,对照组采用脱细胞同种异体神经移植物修复大鼠坐骨神经1cm 缺损,实验组于术后第2天开始给予小剂量超短波治疗,7min/天,1次/天,直至取材,后两组分别于术后第2、4、8、12周时检测患侧L4脊髓内BDNF及CGRP蛋白表达,术后第12周时行患侧胫前肌湿重比率及再生神经电生理检测。结果：①对照组术后第2周时患侧L4脊髓内BDNF表达已开始增高,第4周时达高峰,第8周后逐渐降低,第12周时仍显著高于正常对照组,与对照组相比,实验组只有在第8周时BDNF蛋白表达明显高于对照组（P<0.05）,其余各时间点差异无显著性意义。②对照组患侧L4脊髓内CGRP蛋白表达与BDNF表达趋势基本一致,实验组在术后各时间点脊髓内CGRP蛋白表达均明显高于对照组（P<0.05）。③与对照组相比,术后第12周时实验组再生神经传导速度加快,波幅升高,潜伏时缩短,胫前肌湿重比率明显增加（P<0.05）。结论：小剂量超短波可以促进大鼠周围神经缺损修复术后神经髓鞘及轴索的再生,此作用可能与小剂量超短波上调患侧脊髓内CGRP蛋白的表达,延长BDNF表达的高峰持续时间有关。     

Three‐dimensional printed polycaprolactone‐based scaffolds provide an advantageous environment for osteogenic differentiation of human adipose‐derived stem cells

The capacity of bone grafts to repair critical size defects can be greatly enhanced by the delivery of mesenchymal stem cells (MSCs). Adipose tissue is considered the most effective source of MSCs (ADSCs); however, the efficiency of bone regeneration using undifferentiated ADSCs is low. Therefore, this study proposes scaffolds based on polycaprolactone (PCL), which is widely considered a suitable MSC delivery system, were used as a three‐dimensional (3D) culture environment promoting osteogenic differentiation of ADSCs. PCL scaffolds enriched with 5% tricalcium phosphate (TCP) were used. Human ADSCs were cultured in osteogenic medium both on the scaffolds and in 2D culture. Cell viability and osteogenic differentiation were tested at various time points for 42 days. The expression of RUNX2, collagen I, alkaline phosphatase, osteonectin and osteocalcin, measured by real‐time polymerase chain reaction was significantly upregulated in 3D culture. Production of osteocalcin, a specific marker of terminally differentiated osteoblasts, was significantly higher in 3D cultures than in 2D cultures, as confirmed by western blot and immunostaining, and accompanied by earlier and enhanced mineralization. Subcutaneous implantation into immunodeficient mice was used for in vivo observations. Immunohistological and micro‐computed tomography analysis revealed ADSC survival and activity toward extracellular production after 4 and 12 weeks, although heterotopic osteogenesis was not confirmed – probably resulting from insufficient availability of Ca/P ions. Additionally, TCP did not contribute to the upregulation of differentiation on the scaffolds in culture, and we postulate that the 3D architecture is a critical factor and provides a useful environment for prior‐to‐implantation osteogenic differentiation of ADSCs. Copyright © 2016 John Wiley & Sons, Ltd.     

Regenerative effect of adipose tissue‐derived stem cells transplantation using nerve conduit therapy on sciatic nerve injury in rats

This study proposed a biodegradable GGT nerve conduit containing genipin crosslinked gelatin annexed with tricalcium phosphate (TCP) ceramic particles for the regeneration of peripheral nerves. Cytotoxicity tests revealed that GGT‐extracts were non‐toxic and promoted proliferation and neuronal differentiation in the induction of stem cells (i‐ASCs) derived from adipose tissue. Furthermore, the study confirmed the effectiveness of a GGT/i‐ASCs nerve conduit as a guidance channel in the repair of a 10‐mm gap in the sciatic nerve of rats. At eight weeks post‐implantation, walking track analysis showed a significantly higher sciatic function index (SFI) (P < 0.05) in the GGT/i‐ASC group than in the autograft group. Furthermore, the mean recovery index of compound muscle action potential (CMAP) differed significantly between GGT/i‐ASCs and autograft groups (P < 0.05), both of which were significantly superior to the GGT group (P < 0.05). No severe inflammatory reaction in the peripheral nerve tissue at the site of implantation was observed in either group. Histological observation and immunohistochemistry revealed that the morphology and distribution patterns of nerve fibers in the GGT/i‐ASCs nerve conduits were similar to those of the autografts. These promising results achieved through a combination of regenerative cells and GGT nerve conduits suggest the potential value in the future development of clinical applications for the treatment of peripheral nerve injury. Copyright © 2012 John Wiley & Sons, Ltd.     

Characterization of endothelial-like cells derived from human mesenchymal stem cells

BACKGROUND: Blood-derived endothelial progenitor cells (EPC) have been used to treat ischemic disease. However, the number of EPC that can be obtained from adult blood is limited. OBJECTIVE: To characterize endothelial-like cells obtained from human bone marrow and determine their ability to stimulate new blood vessel formation in vivo. METHODS: Mononuclear cells (MNC) were isolated from human bone marrow or umbilical cord blood and cultured in endothelial growth medium (EGM-2). Mesenchymal stem cells (MSC) were isolated from bone marrow and induced to differentiate into endothelial-like cells (MSCE), or adipocytes or osteocytes by growth in EGM-2, adipogenic or osteogenic medium. RESULTS: Cells obtained by culturing bone marrow MNC in EGM-2 formed cord- or tube-like structures when grown on Matrigel(TM) and expressed several endothelial marker proteins. However, cell morphology and the profile of endothelial marker protein expression were different from those of cord blood-derived EPC (cbEPC). Cells with a similar phenotype were obtained by differentiation of MSC into MSCE, which was accompanied by an increase of endothelial marker proteins and a diminished capacity to differentiate into adipocytes. Subcutaneous implantation of MSCE in collagen plugs in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice resulted in formation of functional blood vessels that had incorporated the MSCE. CONCLUSIONS: Our results show that MSCE and cbEPC are different cell types. The formation of functional blood vessels by MSCE, combined with high yields and a reduced capacity to differentiate into other cell types compared with MSC, makes these cells potentially useful for autologous therapy of ischemic disease.     

移植脑源性神经营养因子转染的骨髓间充质干细胞对AD大鼠的影响

目的:研究移植脑源性神经营养因子(BDNF)转染的骨髓间充质干细胞(MSCs)对阿尔茨海默病(AD)大鼠N-甲基-D-天冬氨酸受体1(NMDAR1)及认知功能的影响。方法:将50只SD大鼠随机平均分为正常对照组、假手术组、AD组、MSCs组、BDNF组。正常对照组不予任何处理,其他4组制备AD模型;MSCs组和BDNF组于制模第11天分别注入10μL(约5×106)MSCs或pIRESneo-EGFP-BDNF转染的MSCs。采用Morris水迷宫测试大鼠学习记忆能力,免疫组化染色法测定NMDAR1水平。结果:MSCs组、BDNF组与AD组比较平均逃避潜伏期(AEL)明显缩短、跨平台次数显著增加;BDNF组较MSCs组改善更显著(P<0.01),NMDAR1水平更高(P<0.01)。结论:BDNF转染骨髓MSCs对AD大鼠的认知有改善作用,且促进海马区NMDAR1的表达。     

Accelerated outgrowth in cross‐facial nerve grafts wrapped with adipose‐derived stem cell sheets

In this study, we devised a novel cross‐facial nerve grafting (CFNG) procedure using an autologous nerve graft wrapped in an adipose‐derived stem cell (ADSC) sheet that was formed on a temperature‐responsive dish and examined its therapeutic effect in a rat model of facial palsy. The rat model of facial paralysis was prepared by ligating and transecting the main trunk of the left facial nerve. The sciatic nerve was used for CFNG, connecting the marginal mandibular branch of the left facial nerve and the marginal mandibular branch of the right facial nerve. CFNG alone, CFNG coated with an ADSC suspension, and CFNG wrapped in an ADSC sheet were transplanted in eight rats each, designated the CFNG, suspension, and sheet group, respectively. Nerve regeneration was compared histologically and physiologically. The time to reinnervation, assessed by a facial palsy scoring system, was significantly shorter in the sheet group than in the other two groups. Evoked compound electromyography showed a significantly higher amplitude in the sheet group (4.2 ± 1.3 mV) than in the suspension (1.7 ± 1.2 mV) or CFNG group (1.6 ± 0.8 mV; p < .01). Toluidine blue staining showed that the number of myelinated fibers was significantly higher in the sheet group (2,450 ± 687) than in the suspension (1,645 ± 659) or CFNG group (1,049 ± 307; p < .05). CFNG in combination with ADSC sheets, prepared using temperature‐responsive dishes, promoted axonal outgrowth in autologous nerve grafts and reduced the time to reinnervation.