Engineered functional organs or tissues,created with autologous somatic cells and seeded on biodegradable or hydrogel scaffolds,have been developed for use in individualswith tissue damage suffered fromcongenital disorders,infection,irradiation,or cancer.However,in those patients,abnormal cells obtained by biopsy fromthe compromised tissue could potentially contaminate the engineered tissues.Thus,an alternative cell source for construction of the neo-organ or functional recovery of the injured or diseased tissues would be useful.Recently,we have found stem cells existing in the urine.These cells are highly expandable,and have self-renewal capacity,paracrine properties,and multi-differentiation potential.As a novel cell source,urine-derived stem cells(USCs)provide advantages for cell therapy and tissue engineering applications in regeneration of various tissues,particularly in the genitourinary tract,because they originate from the urinary tract system.Importantly,USCs can be obtained via a non-invasive,simple,and low-cost approach and induced with high efficiency to differentiate into three dermal cell lineages.
目的:研究抗雄激素受体氟他胺(Flutamide,Flu)孕中、晚期诱导子代雄鼠发生隐睾,及其病理组织学损害,探讨其不育的病理学基础,为临床治疗隐睾所致非梗阻性不育提供依据。方法:将20只SD(sprague dawley)孕鼠随机分为Flu隐睾组(n=10)、正常对照组(n=10),Flu隐睾组于妊娠12~21 d给予Flu 25 mg/(kg·d)皮下注射,正常对照组不予任何处理,取各组出生后第60天(postnatal day 60,PND60)的雄性SD大鼠睾丸组织(Flu隐睾组只取隐睾睾丸),HE染色观察睾丸组织形态学的差异,透射电镜观察睾丸支持细胞间的紧密连接结构,TUNEL凋亡染色检测生精细胞凋亡情况,取附睾尾进行精子计数及形态观测,免疫组织化学和Western blot检测睾丸组织中生殖细胞增殖分化指标视黄酸刺激因子8(stimulated by retinoic acid gene 8,Stra8)、联会复合体蛋白3(synaptonemal complex protein 3,SCP3)的表达,采用Q-PCR检测Stra8基因水平。结果:收集正常对照组正常睾丸30只与Flu隐睾组隐睾睾丸22只,HE染色显示隐睾睾丸管腔明显缩窄、生精细胞发育迟缓且排列紊乱、管腔中央无精子形成;TUNEL凋亡检测证实隐睾组有大量生精细胞发生凋亡,精子计数结果为隐睾组(1.99±0.13)×108个/m L,远低于正常对照组[(5.53±0.17)×108个/m L,P=0.000];透射电镜(transmission electron microscope,TEM)可观察到SD大鼠隐睾支持细胞间的紧密连接结构疏松;免疫组织化学显示Stra8在隐睾组织中表达较正常对照组少,SCP 3在正常对照组睾丸生精小管的各级生精细胞胞核中均有表达,而在隐睾组睾丸中表达很弱;Western blot结果显示,Flu隐睾组Stra8蛋白表达量(0.34±0.05)明显低于正常对照组(0.96±0.09),P=0.002;Flu隐睾组SCP3蛋白表达量(0.39±0.03)也明显低于正常对照组(0.97±0.07),P=0.001。Q-PCR结果显示,Flu组SD大鼠睾丸组织内Stra8 m RNA的表达(0.765±0.015)较正常对照组(1.00±0.01)低,P=0.01。结论:Flu诱导的SD大鼠隐睾模�
