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1.
孙锡金  胡志奇  易斌 《中国美容医学》2006,15(8):886-888,i0001
目的:寻求适合于毛囊组织工程的大规模高效快速分离培养人头皮毛乳头细胞的方法。方法:将含有毛囊中下部的皮下组织剪碎,加入胶原酶Ⅰ消化2h,悬液经400目筛网过滤,收集被筛网截留的毛乳头进行培养,计算其贴壁率,绘制细胞生长曲线,并与显微解剖法相比较。结果:用这种方法成功分离培养了人头皮毛乳头细胞。结论:消化过滤法是一种能够大规模高效、快速分离培养人头皮毛乳头细胞的方法。  相似文献   

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改良一步酶消化法分离培养人头皮毛乳头细胞的实验研究   总被引:1,自引:0,他引:1  
目的进一步简化人头皮毛乳头的分离方法,实现高纯度毛乳头细胞的体外扩增。方法将含有毛球部的头皮皮下组织剪成肉泥状,胶原酶Ⅰ(2mg/ml)37℃消化2~3h后,加入10倍体积的DMEM,以吸管反复吹打消化液使毛乳头全部游离出来,在显微镜下用微量移液器逐个收集表面无杂质黏附的毛乳头。结果“改良一步酶消化法”分离毛乳头所需的操作时间进一步缩短,劳动强度降低;所得毛乳头纯度高,贴壁率达到99%,其细胞迁出加速。结论“改良一步酶消化法”是一种高效、快速分离培养人头皮毛乳头细胞的方法。  相似文献   

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目的进一步简化人头皮毛乳头的分离方法,实现高纯度毛乳头细胞的体外扩增.方法将含有毛球部的头皮皮下组织剪成肉泥状,胶原酶Ⅰ(2 mg/ml) 37 ℃消化2~3 h后,加入10倍体积的DMEM,以吸管反复吹打消化液使毛乳头全部游离出来,在显微镜下用微量移液器逐个收集表面无杂质黏附的毛乳头.结果 "改良一步酶消化法" 分离毛乳头所需的操作时间进一步缩短,劳动强度降低;所得毛乳头纯度高,贴壁率达到99%,其细胞迁出加速.结论 "改良一步酶消化法"是一种高效、快速分离培养人头皮毛乳头细胞的方法.  相似文献   

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目的 研究不同血清浓度的培养基对毛乳头细胞生长速度及细胞状态的影响.方法 改良一步酶消化法分离培养人头皮毛乳头细胞,分别以无血清培养基及10%、15%等不同浓度小牛血清培养基,培养第3代毛乳头细胞,倒置相差显微镜下观察细胞生长状态,细胞消化后,计数并绘制生长曲线.结果 在无血清培养基培养条件下,毛乳头细胞传代后贴壁率较低.贴壁后细胞未见明显增殖;传代后前8 d,15%和20%血清培养基培养的毛乳头细胞生长速度,明显快于无血清培养基组和10%血清培养基组(P<0.05);传代第10天后,15%和20%血清培养基中毛乳头细胞生长速度差异无显著的统计学意义.结论 细胞生长速度与血清浓度密切相关.  相似文献   

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微囊化人头皮毛乳头细胞体外培养及异种移植   总被引:1,自引:1,他引:0  
目的探讨微囊化人头皮毛乳头细胞体外培养及异种移植的可行性,并对海藻酸钠-多聚赖氨酸-海藻酸钠(alginate-polylysine-alginate,APA)微囊与海藻酸钠-BaCi2(barium-alginate,BA)微囊的物理、生物性能进行评价.方法采用“一步酶消化法”分离、培养人头皮毛乳头细胞,分别用APA微囊与BA微囊包裹.对两种微囊的生物相容性、机械强度、免疫隔离效果及微囊内细胞活性进行比较.结果APA微囊生物相容性优于BA微囊(P<0.01),但机械强度低于BA微囊(P<0.01);成囊后短期BA微囊内细胞活性高于APA微囊(P<0.01),但APA微囊内细胞活性增高较快(P<0.05).在微囊完整、表面无纤维化时,两种微囊均可起到良好的免疫隔离作用.结论微囊化人头皮毛乳头细胞可在体外及异种体内培养.综合评价APA微囊与BA微囊的利弊,在不同情况下选择不同的成囊方式是必要的.  相似文献   

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目的 寻求一种洁净且无须显微镜下操作的毛乳头分离方法 ,培养高纯度的毛乳头细胞.方法 中性蛋白酶消化头皮组织,将含有毛囊的皮下脂肪层剪下切碎,并用Ⅰ型胶原酶消化,之后通过多次离心的方法 获得毛乳头;对培养出的毛乳头细胞进行形态学观察,并检测毛乳头细胞的标志物alfa-SMA和碱性磷酸酶的表达情况.结果 通过新型的两步酶消化法可获得洁净的毛乳头,培养出的毛乳头细胞标志物检测阳性.结论 新型的两步酶消化法是一种高效低劳动强度的毛乳头分离方法.  相似文献   

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目的观察人头皮毛乳头细胞微囊(人工毛乳头)异种移植诱导大鼠足垫毛囊形成的能力。方法以海藻酸钠-多聚赖氨酸-海藻酸钠(alginate-polylysine-alginate,APA)微囊包裹分离培养的毛乳头细胞;对体外培养1、4周的毛乳头细胞微囊及无APA的微囊对照组行组织学观察;取培养4周的毛乳头细胞微囊移植至大鼠足垫皮下,6周后取材行组织学检查。结果毛乳头细胞微囊体外培养1周后,毛乳头细胞周围出现细胞外基质;4周后,囊中形成“类毛乳头样结构”;人头皮毛乳头细胞微囊移植至大鼠足垫6周后,移植部位及其周围皮下有大量毛囊及皮脂腺结构形成。结论人工毛乳头诱导并参与了无毛区域新生毛囊及皮脂腺的组织构成。  相似文献   

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人工毛乳头异种移植诱导大鼠足垫毛囊形成   总被引:3,自引:1,他引:2  
目的观察人头皮毛乳头细胞微囊(人工毛乳头)异种移植诱导大鼠足垫毛囊形成的能力。方法以海藻酸钠-多聚赖氨酸-海藻酸钠(alginate- polylysine - alginate, APA)微囊包裹分离培养的毛乳头细胞;对体外培养1、4周的毛乳头细胞微囊及无APA的微囊对照组行组织学观察;取培养4周的毛乳头细胞微囊移植至大鼠足垫皮下,6周后取材行组织学检查。结果毛乳头细胞微囊体外培养1周后,毛乳头细胞周围出现细胞外基质;4周后,囊中形成“类毛乳头样结构”;人头皮毛乳头细胞微囊移植至大鼠足垫6周后,移植部位及其周围皮下有大量毛囊及皮脂腺结构形成。结论人工毛乳头诱导并参与了无毛区域新生毛囊及皮脂腺的组织构成。  相似文献   

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目的毛乳头细胞被广泛用于组织工程毛囊和皮肤构建。探讨用含血清的角质形成细胞培养基(keratinocyte medium,KM)和普通培养基(normal medium,NM)培养毛乳头细胞的生物学特性差异,以及KM培养的毛乳头细胞是否可用于组织工程毛囊研究。方法采用两步酶消化法从除皱术时切下的头皮标本中分离培养毛乳头,接种后分别采用KM和NM培养毛乳头细胞并传代。记录毛乳头最早贴壁时间及细胞迁出时间,接种5d时统计毛乳头贴壁率。倒置显微镜下观察两组细胞形态学变化,计数第3代细胞每皿聚集体个数;记录细胞最大传代次数;细胞融合成膜后对消化的细胞膜片行HE染色观察。取第3代细胞行MTT法检测细胞增殖;免疫荧光染色和ALP染色观察α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和ALP在两组细胞中的表达,并统计ALP蓝色阳性区域面积。结果KM组毛乳头最早贴壁时间及细胞迁出时间分别为24h及64h,早于NM组的48h及80h。5d时KM组及NM组毛乳头贴壁率分别为54.17%及36.78%。NM组细胞体积较KM组大。NM组第3代细胞每皿形成(3.06±1.12)个聚集体,KM组每皿形成(9.25±1.73)个聚集体,两组比较差异有统计学意义(P0.05)。KM组细胞融合后细胞膜片行HE染色可见细胞呈多层性;NM组细胞不能多层性生长,未行HE染色观察。NM组细胞最多可传7代;KM组最多可传15代。MTT检测显示KM组细胞增殖迅速,7d后KM组吸光度值明显高于NM组,差异有统计学意义(P0.05)。免疫荧光染色显示,两组第3代细胞α-SMA均为阳性。ALP染色显示,NM组第6代细胞中仅少量细胞表达ALP,阳性区域面积为(987±146)μm2;KM组第14代细胞ALP染色仍为阳性,阳性区域面积为(8757±558)μm2;两组比较差异有统计学意义(P0.05)。结论毛乳头细胞在含血清的KM中增殖迅速,聚集生长明显,传代次数增多;用含血清KM培养毛乳头细胞用于组织工程毛囊构建是可行的。  相似文献   

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微囊化人头皮毛乳头细胞移植诱导毛发形成   总被引:5,自引:0,他引:5  
目的探索微囊化人头皮毛乳头细胞(以下简称毛乳头细胞)对裸鼠背部皮肤毛囊形成的诱导作用。方法胶原酶消化法体外分离、培养毛乳头细胞,再将毛乳头细胞以海藻酸钠-多聚赖氨酸-海藻酸钠(alginate-polylysine—alginate,APA)微囊包裹,以胶原凝胶作为载体,植入裸鼠背部皮下;移植空囊、游离毛乳头细胞各作为对照。观察移植部位毛发生长情况,利用组织学方法观测所形成的毛囊结构。结果微囊化毛乳头细胞皮下移植4周后,裸鼠背部移植区有白色、浓密、分布均匀的毛发长出。局部组织切片见大量发育完整的毛囊结构;而空囊及游离毛乳头细胞移植均未能诱导出上述现象。结论聚集性生长的微囊化毛乳头细胞能够保持诱导皮肤毛囊形成的作用,且这种作用无种属特异性。  相似文献   

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BackgroundAbsenteeism is costly, yet evidence suggests that presenteeism—illness-related reduced productivity at work—is costlier. We quantified employed patients’ presenteeism and absenteeism before and after total joint arthroplasty (TJA).MethodsWe measured presenteeism (0-100 scale, 100 full performance) and absenteeism using the World Health Organization’s Health and Work Performance Questionnaire before and after TJA among a convenience sample of employed patients. We captured detailed information about employment and job characteristics and evaluated how and among whom presenteeism and absenteeism improved.ResultsIn total, 636 primary, unilateral TJA patients responded to an enrollment email, confirmed employment, and completed a preoperative survey (mean age: 62.1 years, 55.3% women). Full at-work performance was reported by 19.7%. Among 520 (81.8%) who responded to a 1-year follow-up, 473 (91.0%) were still employed, and 461 (88.7%) had resumed working. Among patients reporting at baseline and 1 year, average at-work performance improved from 80.7 to 89.4. A Wilcoxon signed-rank test indicated that postoperative performance was significantly higher than preoperative performance (P < .0001). The percentage of patients who reported full at-work performance increased from 20.9% to 36.8% (delta = 15.9%, 95% confidence interval = [10.0%, 21.9%], P < .0001). Presenteeism gains were concentrated among patients who reported declining work performance leading up to surgery. Average changes in absences were relatively small. Combined, the average monthly value lost by employers to presenteeism declined from 15.3% to 8.3% and to absenteeism from 16.9% to 15.5% (ie, mitigated loss of 8.4% of monthly value).ConclusionAmong employed patients before TJA, presenteeism and absenteeism were similarly costly. After, employed patients reported increased performance, concentrated among those with declining performance leading up to surgery.  相似文献   

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As well for optimized emergency management in individual cases as for optimized mass medicine in disaster management, the principle of the medical doctors approaching the patient directly and timely, even close to the site of the incident, is a long-standing marker for quality of care and patient survival in Germany. Professional rescue and emergency forces, including medical services, are the “Golden Standard” of emergency management systems. Regulative laws, proper organization of resources, equipment, training and adequate delivery of medical measures are key factors in systematic approaches to manage emergencies and disasters alike and thus save lives. During disasters command, communication, coordination and cooperation are essential to cope with extreme situations, even more so in a globalized world. In this article, we describe the major historical milestones, the current state of the German system in emergency and disaster management and its integration into the broader European approach.  相似文献   

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Bone defects related to osteoporosis develop with increasing age and differ between males and females. It is currently thought that the bone remodeling process is supervised by osteocytes in a strain-dependent manner. We have shown an altered response of osteocytes from osteoporotic patients to mechanical loading, and osteocyte density is reduced in osteoporotic patients, which might relate to imperfect bone remodeling, leading to lack of bone mass and strength. Hence, information on osteocyte density will contribute to a better understanding of bone biology in males and females and to the assessment of osteoporosis. Osteocyte density as well as conventional histomorphometric parameters of trabecular bone were determined in cancellous iliac crest bone of healthy postmenopausal women and men and of osteoporotic women and men. Osteocyte density was higher in healthy females than in healthy males and lower in osteoporotic females than in healthy females. Bone mass was reduced in osteoporotic patients, both male and female. In females, trabecular number was reduced, whereas in males, trabecular thickness was reduced and eroded surface was increased. There were no correlations between the parameter groups bone architecture, bone formation, bone resorption, and osteocyte density. These results are consistent with impaired osteoblast function in osteoporotic patients and with a different mechanism of bone loss between men and women, in which osteocyte density might play a role. The reduced osteocyte numbers in female osteoporotic patients might relate to imperfect bone remodeling leading to lack of bone mass and strength. M. G. Mullender and S. D. Tan contributed equally to this work.  相似文献   

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目的探讨肝内胆管囊腺瘤和囊腺癌的CT、MRI和病理特点。方法回顾性分析经手术病理证实的6例肝内胆管囊腺瘤和2例肝内胆管囊腺癌的影像及临床病理资料,将病变的影像表现与其病理大体形态及组织学表现作对照分析。结果6例肝内胆管囊腺瘤,女4例、男2例;2例肝内胆管囊腺癌均为女性病人;8例病人平均年龄55岁。所有病灶均表现为多房囊性肿块,肿瘤囊腔各分房内常为多种液体成分,在CT上可表现为不同密度、在MRI上可表现为不同信号强度。囊内出现多发大小不等的壁结节在胆管囊腺癌内更常见,囊内有分隔但无壁结节只见于胆管囊腺瘤。在7例CT扫描中,4例胆管囊腺瘤和1例胆管囊腺癌可见囊壁或分隔上钙化,囊壁、囊内分隔及囊内结节均为轻、中度延迟增强。肿瘤中出现卵巢样间质见于3例胆管囊腺瘤和1例胆管囊腺癌,且均为女性病人。结论肝内胆管囊腺瘤和囊腺癌是肝脏不常见的囊性肿瘤,影像上多房、囊内有分隔且各分房囊内密度或信号不一致,高度提示肝内胆管囊腺瘤或囊腺癌的诊断,如囊内伴有多发大小不等的结节,则进一步提示囊腺癌的可能。但影像学表现不能区分肿瘤中有无卵巢样间质。  相似文献   

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