基于脂肪干细胞的皮肤创面汗腺再生机制研究

A large area of burn wound surface has resulted in loss of sweat gland,which has seriously influenced the function of perspiration and caused patients not to regulate their body-temperature effectively.As to burn patients,lack of autogenic epidermal stem cells has been one of the biggest problem in promoting sweat gland regeneration.Due to lack of autogeneic epidermal stem cells,the other types of stem cells have been explored to improve the level of sweat gland regeneration.However,no excellent kind of stem cells has been found until now.The application of embryo stem cells has faced the challenge of ethics,and the large quantity of bone marrow stem cells has't been easily obtained in burn patients.Thereforce,the important problem has been to find ideal stem cells and in turn effectively induce them to differentiate toward sweat gland epithelial cells..Recent studies have already found that autogeneic adipose derived stem cells(ADSCs) have the potential ability to transdifferentiation across multiblasto -derms,which make us logically hypothesize that we can induce transdifferentiation of ADSCs toward sweat gland epithelial cells and in turn improve the level of sweat gland regeneration on burn wound surface by the application of ADSCs..Our research will include in vitro study and in vivo study.In vitro study,ADSCs will be obtained from fat liposuction.After the assessment of surface antigen concern -ing ADSCs through flow cytometry and the evaluation of multipotential differentia -tion of ADSCs,EDA-A1 gene will be novelly transfected into ADSCs through adenovirus vector,then ADSCs will be cocultured with EGF、homogenate of sweat gland tissue in order to induce their transdifferentiation toward sweat gland epithelial cells.The optimal condition concerning transdifferentiation of ADSCs toward sweat gland epithelial cells will be established and obtained sweat gland epithelical cells..In vivo study,The burn animal model of athymic mice,which has been widely proven by academic circles,will be set up.Then,the sweat gland epithelial cells or EDA-A1 transfected ADSCs with different inducing factors will be transplanted into the skin wound surface of athymic mice.The biological safety and the influence of the different cells on sweat gland regeneration will be evaluated..This research will elucidate the feasibility and optimal style of the application of ADSCs in promoting sweat gland regeneration.On basis of the study,one novel method will be found in improving the level of sweat gland regeneration,which has the importantly theoretical and potentially clinical significance.

大面积皮肤创面常伴汗腺缺失,影响体温调节,带给患者无尽痛苦,因此促进汗腺再生意义重大。自身表皮干细胞缺乏是汗腺再生的瓶颈,目前应用胚胎或骨髓干细胞促进汗腺再生存在伦理学困扰和获取困难两大障碍无法逾越;因此寻找理想干细胞并诱导其向汗腺上皮细胞分化成为汗腺再生的关键。我们前期证实脂肪干细胞经特定诱导后有向汗腺上皮细胞分化的趋势,据此提出科学假设:"诱导脂肪干细胞横向分化为汗腺上皮细胞并促进汗腺再生"。本研究首次把汗腺发育相关基因EDA-A1导入脂肪干细胞,然后在EGF、汗腺组织匀浆等条件下诱导,建立脂肪干细胞向汗腺上皮细胞分化的诱导体系;然后把体外实验所获汗腺上皮细胞，或EDA-A1转染的脂肪干细胞联合不同诱导因子移植到裸鼠创面,并评价上述细胞移植的生物安全性及其对汗腺再生的影响。本项目将阐明脂肪干细胞促进汗腺再生的可行性及其最佳作用方式,消除制约汗腺再生的瓶颈,为汗腺再生提供新途径和理论基础

大面积皮肤缺损造成汗腺缺失影响体温调节功能，给患者带来巨大的痛苦，因此促进汗腺再生意义重大。脂肪干细胞具有跨胚层分化能力且取材容易，不受伦理学困扰而成为汗腺再生的种子细胞之一。本研究成功分离了人脂肪干细胞并完成传代培养和鉴定。通过构建腺病毒表达载体转染EDA-A基因至脂肪干细胞，然后通过EGF等生长因子，联合汗腺组织匀浆与汗腺上皮细胞Transwell共培养等措施诱导脂肪干细胞向汗腺上皮细胞分化。通过免疫荧光及流式细胞技术鉴定细胞表型，结果表明人脂肪干细胞通过诱导可分化为汗腺上皮样细胞。继而制备动物模型初步探索了应用人脂肪干细胞来源汗腺上皮细胞促进汗腺组织再生，取得了已经的经验和进展。本研究证实了人脂肪干细胞促进汗腺再生的可行性，为汗腺再生提供了新的手段，具有十分诱人的研究前景和临床转化潜能。本研究已发表论文7篇，其中SCI收录4篇，培养硕士研究生8名，博士研究生1名。