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2.
人脂肪干细胞结合微载体在生物反应器中向软骨细胞分化 总被引:3,自引:0,他引:3
[目的]探索在旋转生物反应器内,应用微载体技术快速扩增并向软骨分化人脂肪干细胞。[方法]将人脂肪干细胞结合Cytodex3微载体在旋转的生物反应器(RCSS)内进行动态培养,应用倒置显微镜和扫描电镜对微载体表面的脂肪干细胞进行动态观察,并对收获的脂肪干细胞进行Safran in-O、tolu id ine b lue染色等组织化学染色及Ⅱ型胶原的免疫化学染色分析。[结果]脂肪干细胞于24 h内贴附于Cytodex3微载体表面,细胞形态为短梭形,随时间的延长,贴附于微载体的细胞逐渐增多,到培养后期,细胞密度可达最初接种的19倍左右,在微载体上收获的细胞进行番红花O、阿利新蓝染色呈阳性,Ⅱ型胶原染色阳性,均强于对照组。[结论]利用微载体细胞培养技术可简便快速地在体外扩增脂肪干细胞,并成功实现向软骨细胞分化。 相似文献
3.
Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor 总被引:5,自引:0,他引:5
Aizaki H Nagamori S Matsuda M Kawakami H Hashimoto O Ishiko H Kawada M Matsuura T Hasumura S Matsuura Y Suzuki T Miyamura T 《Virology》2003,314(1):16-25
Lack of efficient culture systems for hepatitis C virus (HCV) has been a major obstacle in HCV research. Human liver cells grown in a three-dimensional radial-flow bioreactor were successfully infected following inoculation with plasma from an HCV carrier. Subsequent detection of increased HCV RNA suggested viral replication. Furthermore, transfection of HCV RNA transcribed from full-length cDNA also resulted in the production and release of HCV virions into supernatant. Infectivity was shown by successful secondary passage to a new culture. Introduction of mutations in RNA helicase and polymerase regions of HCV cDNA abolished virus replication, indicating that reverse genetics of this system is possible. The ability to replicate and detect the extracellular release of HCV might provide clues with regard to the persistent nature of HCV infection. It will also accelerate research into the pathogenicity of HCV, as well as the development of prophylactic agents and new therapy. 相似文献
4.
静态与动态下脱细胞猪主动脉瓣种植人成纤维细胞初步研究 总被引:1,自引:0,他引:1
目的 探索在脱细胞猪主动脉瓣膜材料上种植人成纤维细胞的方法与效果;材料和方法 采用在脱细胞猪主动脉瓣膜材料上静态与动态种植人成纤维细胞,并对其结果进行比较研究。结果 静态种植不能在猪主动脉瓣叶的两侧形成完整的单层人成纤维细胞融合层;而经过动态种植,在一定条件下可以在猪主动脉瓣叶两侧形成完整的单层人成纤维细胞融合层;结论 动态种植较之静态种植易达到形成完整均匀的细胞种植效果。 相似文献
5.
软骨组织工程中力学因素的影响及应用 总被引:1,自引:0,他引:1
力学因素是软骨组织工程中的重要影响因素之一。近年来的研究表明,力学作用可以刺激细胞因子及激素的分泌,改变三维支架上培养的软骨细胞的新陈代谢,从而促进软骨组织的生长与重建。目前已经有诸多关于体外构建软骨组织的报道,但对于其中的力学因素的影响(包括力学因素对软骨细胞增殖的促进及力学刺激的传导机制等)还没有完全认识。就以上几方面做一综述,并简单介绍生物反应器在软骨组织工程中的应用。 相似文献
6.
Development of a novel pulsatile bioreactor for tissue culture 总被引:1,自引:1,他引:1
Yos S. Morsi William W. Yang Amal Owida Cynthia S. Wong 《Journal of artificial organs》2007,10(2):109-114
The construction of tissue-engineered parts such as heart valves and arteries requires more than just the seeding of cells
onto a biocompatible/biodegradable polymeric scaffold. It is essential that the functionality and mechanical integrity of
the cell-seeded scaffold be investigated in vitro prior to in vivo implantation. The correct hemodynamic conditioning would
lead to the development of tissues with enhanced mechanical strength and cell viability. Therefore, a bioreactor that can
simulate physiological conditions would play an important role in the preparation of tissue-engineered constructs. In this
article, we present and discuss the design concepts and criteria, as well as the development, of a multifunctional bioreactor
for tissue culture in vitro. The system developed is compact and easily housed in an incubator to maintain sterility of the
construct. Moreover, the proposed bioreactor, in addition to mimicking in vivo conditions, is highly flexible, allowing different
types of constructs to be exposed to various physiological flow conditions. Initial verification of the hemodynamic parameters
using Laser doppler anemometry indicated that the bioreactor performed well and produced the correct physiological conditions. 相似文献
7.
《Expert opinion on biological therapy》2013,13(5):677-682
Due to its accessibility, size and contact with the blood circulation, the skin is an attractive target for somatic gene therapy. Permanent cutaneous expression can be achieved by genetic manipulation of epidermal keratinocytes ex vivo followed by transplantation or by local injection of viral vectors. Furthermore, progress is being made to develop topical gene transfer methods leading to permanent gene expression. There is experimental evidence showing that genetically engineered skin can produce and secrete medically relevant proteins to the circulation and also produce enzymes that can clear metabolites accumulating in various diseases. Thus, cutaneous gene transfer approaches may be relevant not only for local skin diseases, but also for certain systemic disorders. 相似文献
8.
Deformation strain is the main physical driver for skeletal precursors to undergo osteogenesis in earlier stages of osteogenic cell maturation
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Anna Finne‐Wistrand Melanie Krug Thomas Schwarz Franz Jakob Heike Walles Jan Hansmann 《Journal of tissue engineering and regenerative medicine》2018,12(3):e1474-e1479
Mesenchymal stem cells play a major role during bone remodelling and are thus of high interest for tissue engineering and regenerative medicine applications. Mechanical stimuli, that is, deformation strain and interstitial fluid‐flow‐induced shear stress, promote osteogenic lineage commitment. However, the predominant physical stimulus that drives early osteogenic cell maturation is not clearly identified. The evaluation of each stimulus is challenging, as deformation and fluid‐flow‐induced shear stress interdepend. In this study, we developed a bioreactor that was used to culture mesenchymal stem cells harbouring a strain‐responsive AP‐1 luciferase reporter construct, on porous scaffolds. In addition to the reporter, mineralization and vitality of the cells was investigated by alizarin red staining and 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide. Quantification of the expression of genes associated to bone regeneration and bone remodelling was used to confirm alizarin red measurements. Controlled perfusion and deformation of the 3‐dimensional scaffold facilitated the alteration of the expression of osteogenic markers, luciferase activity, and calcification. To isolate the specific impact of scaffold deformation, a computational model was developed to derive a perfusion flow profile that results in dynamic shear stress conditions present in periodically loaded scaffolds. In comparison to actually deformed scaffolds, a lower expression of all measured readout parameters indicated that deformation strain is the predominant stimulus for skeletal precursors to undergo osteogenesis in earlier stages of osteogenic cell maturation. 相似文献
9.
Mandana Haack-Sørensen Morten Juhl Bjarke Follin Rebekka Harary Søndergaard Maria Kirchhoff Jens Kastrup 《Scandinavian journal of clinical and laboratory investigation》2018,78(4):293-300
In vitro expanded adipose-derived stromal cells (ASCs) are a useful resource for tissue regeneration. Translation of small-scale autologous cell production into a large-scale, allogeneic production process for clinical applications necessitates well-chosen raw materials and cell culture platform. We compare the use of clinical-grade human platelet lysate (hPL) and fetal bovine serum (FBS) as growth supplements for ASC expansion in the automated, closed hollow fibre quantum cell expansion system (bioreactor). Stromal vascular fractions were isolated from human subcutaneous abdominal fat. In average, 95?×?106 cells were suspended in 10% FBS or 5% hPL medium, and loaded into a bioreactor coated with cryoprecipitate. ASCs (P0) were harvested, and 30?×?106 ASCs were reloaded for continued expansion (P1). Feeding rate and time of harvest was guided by metabolic monitoring. Viability, sterility, purity, differentiation capacity, and genomic stability of ASCs P1 were determined. Cultivation of SVF in hPL medium for in average nine days, yielded 546?×?106 ASCs compared to 111?×?106 ASCs, after 17?days in FBS medium. ASCs P1 yields were in average 605?×?106 ASCs (PD [population doublings]: 4.65) after six days in hPL medium, compared to 119?×?106 ASCs (PD: 2.45) in FBS medium, after 21?days. ASCs fulfilled ISCT criteria and demonstrated genomic stability and sterility. The use of hPL as a growth supplement for ASCs expansion in the quantum cell expansion system provides an efficient expansion process compared to the use of FBS, while maintaining cell quality appropriate for clinical use. The described process is an obvious choice for manufacturing of large-scale allogeneic ASC products. 相似文献
10.
Cyclic mechanical stimulation rescues achilles tendon from degeneration in a bioreactor system
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![点击此处可从《Journal of orthopaedic research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Tao Wang Zhen Lin Ming Ni Christine Thien Robert E. Day Bruce Gardiner Jonas Rubenson Thomas B. Kirk David W. Smith Allan Wang David G. Lloyd Yan Wang Qiujian Zheng Ming H. Zheng 《Journal of orthopaedic research》2015,33(12):1888-1896
Physiotherapy is one of the effective treatments for tendinopathy, whereby symptoms are relieved by changing the biomechanical environment of the pathological tendon. However, the underlying mechanism remains unclear. In this study, we first established a model of progressive tendinopathy‐like degeneration in the rabbit Achilles. Following ex vivo loading deprivation culture in a bioreactor system for 6 and 12 days, tendons exhibited progressive degenerative changes, abnormal collagen type III production, increased cell apoptosis, and weakened mechanical properties. When intervention was applied at day 7 for another 6 days by using cyclic tensile mechanical stimulation (6% strain, 0.25 Hz, 8 h/day) in a bioreactor, the pathological changes and mechanical properties were almost restored to levels seen in healthy tendon. Our results indicated that a proper biomechanical environment was able to rescue early‐stage pathological changes by increased collagen type I production, decreased collagen degradation and cell apoptosis. The ex vivo model developed in this study allows systematic study on the effect of mechanical stimulation on tendon biology. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1888–1896, 2015. 相似文献