目的大面积深度烧伤治疗的半数致死面积(LA50)已达98%,但由于伤面切除植皮后无汗液分泌,使患者的生活质量显著下降。目前,由于烧伤治疗技术已趋成熟,存活者日益增多,因而汗腺再生的问题必须解决,而通过干细胞移植诱导再生汗腺是其中的重要方法之一。方法分离培养人骨髓间充质干细胞(MSCs)和人汗腺细胞(SGCs)并进行鉴定。通过共培养方式促进骨髓间充质干细胞向汗腺细胞分化。将具有汗腺表型的干细胞局部移植于裸鼠烫伤脚掌,观察汗腺组织再生修复情况,结合碘-淀粉发汗试验检测汗腺功能以判定治疗效果。在动物实验初步证明转化的汗腺细胞能发挥修复汗腺功能后,选择2例烧伤自愿患者。1例患者双上臂背侧均有相似的烧伤后遗留的瘢痕,经发汗腺试验证明无汗液分泌。抽取其骨髓并切取小块正常皮肤分离培养骨髓间充质干细胞和汗腺细胞进行共培养,诱导骨髓间充质干细胞获得汗腺细胞表型,通过自行设计的方案将该细胞种植于右侧切除瘢痕处。创面愈合后治疗侧行发汗试验、汗液成分分析以及组织学检查。另1例为颏颈挛缩、瘢痕切除解除挛缩的患者,在其1/3的创面上种植诱导的汗腺细胞,2个月后进行发汗试验。结果通过共培养方式培养的骨髓间充质干细胞表达汗腺细胞特异性标志,可获得汗腺细胞表型。发汗试验和形态学观察证实将具有汗腺细胞表型的干细胞移植于裸鼠脚掌的创面后,能显著促进受损汗腺的修复与再生。人体移植试验表明,种植诱导的汗腺细胞后患者创面愈合部位呈现出汗功能,对照部位发汗试验阴性,形态学检测显示移植汗腺的真皮浅层有大量癌胚抗原(CEA)阳性细胞,结构类似正常汗腺组织。汗液成分分析结果显示移植处收集的汗液与正常皮肤处收集的汗液具有类似的pH值、渗透压和化学组分。第2例创?
Background The loss of perspiration after a massive deep burn hampers the survivor to lead a life of high quality, as they are deprived the function of regulating body temperature through perspiration during sultry months. With maturation of science of burn care, the number of survivors is increased, therefore, it is imperative that this problem should be tackled in order to improve their quality of life. Objective To explore the possibility of transdifferentiating bone marrow mesenchymal stem cells (MSCs) into sweat gland cells (SGCs), and implanting the latter into fresh skin wound to generate functional sweat glands. Methods Human bone marrow MSCs and SGCs were isolated from the same patients. They were identified with specific markers, and then co-cultured. The stem cells which subsequently exhibited the phenotype of sweat gland cells were implanted into scald injured paws of nude mice, and regeneration of functioning sweat glands was confirmed by perspiration test (iodine and starch) and histological examination. A male patient bearing almost iden- tical burn scars on the posterior aspect of both arms was enrolled for clinical trial. The scars were first proved to be anhydrotic with iodine and starch test. With patient's written consent, the clinical trial was carried out. Bone marrow MSCs and sweat gland cells were obtained from the patient. After being heat shocked, the SGCs were co-cultured with MSCs. Three days later, the scars of both arms were excised. MSCs having acquired the phenotype of sweat gland cells after co-culture were evenly spread onto the excision wound on the right arm. They were covered with a piece of acellular allogeneic dermis, which was perforated with numerous micropores. On top of the latter, micrografts of autologous origin were transplanted, and the wound was finally covered with a piece of allogeneic skin graft. The wound on the left side was similarly covered, but without transdifferentiated MSCs. After complete healing of the wounds, perspiration test with iodine