本文選題：流體剪切力 + 人骨髓間充質(zhì)干細(xì)胞��； 參考：《第三軍醫(yī)大學(xué)》2005年碩士論文

【摘要】： 生物反應(yīng)器是近年來生物工程方面研究熱點(diǎn)之一,廣泛應(yīng)用于培養(yǎng)細(xì)胞及組織復(fù)合物。灌注式生物反應(yīng)器不僅具有三維動力性培養(yǎng)的優(yōu)點(diǎn),而且其動力性環(huán)境可對種子細(xì)胞產(chǎn)生流體剪切應(yīng)力(fluid shear stress,FSS),因此在骨組織工程研究領(lǐng)域呈現(xiàn)出良好的應(yīng)用前景。骨骼的骨量和正常生理狀態(tài)的維持有賴于對骨骼施加適當(dāng)?shù)膽?yīng)力刺激。研究表明骨骼基質(zhì)形變引起的細(xì)胞外液流動所形成的FSS是作用于骨骼細(xì)胞的主要應(yīng)力。FSS對骨骼細(xì)胞正常生理活動及增殖與分化有著重要的作用。 平行平板流動腔(parallel-plate flow chamber,PPFC)作為一種可以提供較精確液體層流的實(shí)驗(yàn)裝置,常被用于研究細(xì)胞流體力學(xué)。骨髓間充質(zhì)干細(xì)胞(mesenchymal stem cells, MSCs)被認(rèn)為是最有發(fā)展前景的骨組織工程種子細(xì)胞之一。hMSCs存在于骨髓基質(zhì)中,其生理環(huán)境與骨骼細(xì)胞類似,也應(yīng)是應(yīng)力敏感細(xì)胞,推測FSS作用于hMSCs后應(yīng)該產(chǎn)生與骨骼細(xì)胞相似的生物學(xué)效應(yīng)。實(shí)驗(yàn)通過對平行平板流動腔進(jìn)行改良制作,利用其對hMSCs進(jìn)行FSS刺激,通過倒置顯微鏡及透射電鏡觀察細(xì)胞形態(tài)學(xué)改變,細(xì)胞增殖通過MTT法及流式細(xì)胞儀檢測,成骨分化通過堿性磷酸酶和骨鈣素活性測定、堿性磷酸酶染色、鈣結(jié)節(jié)茜素紅染色及四環(huán)素?zé)晒鈽?biāo)記檢測。我們通過觀察FSS刺激hMSCs體外增殖及向成骨方向分化的作用,為進(jìn)一步研究FSS刺激hMSCs體外增殖及分化的機(jī)制、利用灌注式生物反應(yīng)器培養(yǎng)組織工程骨提供理論依據(jù)。主要研究結(jié)果及結(jié)論如下: 一、改良的平行平板流動腔的制備及其優(yōu)點(diǎn)。流動腔以有機(jī)玻璃作為流動腔管道部分,塑料薄膜用來密封流動腔和保持流動腔高度,應(yīng)用塑料細(xì)胞培養(yǎng)瓶底作為細(xì)胞貼附載體,靜脈輸液管作為灌流液循環(huán)管道,以長尾夾固定和密封平行平板流動腔。整個裝置以放射線輻照消毒滅菌。此模型較其它報道的流體剪切力刺激模型制作簡單,成本低廉,操作簡便。實(shí)驗(yàn)證明此系統(tǒng)作為hMSCs流體力學(xué)的研究模型是科學(xué)、可行的。 二、流體剪切力可促進(jìn)人骨髓間充質(zhì)干細(xì)胞增殖。將傳代的第三代hMSCs分為
[Abstract]:Bioreactor is one of the research hotspots in bioengineering in recent years. It is widely used in cultured cells and tissue complexes. Perfusion bioreactor not only has the advantage of three-dimensional dynamic culture, but also its dynamic environment can produce fluid shear stress and shear stress in seed cells, so it has a good application prospect in the field of bone tissue engineering. The maintenance of bone mass and normal physiological state depends on appropriate stress stimulation on bone. The results showed that FSS formed by extracellular fluid flow induced by deformation of bone matrix is the main stress of bone cells. FSs play an important role in normal physiological activities, proliferation and differentiation of bone cells. Parallel-plate flow chamber PPFC (parallel plate flow chamber), as an experimental device which can provide accurate liquid laminar flow, is often used to study cell hydrodynamics. Mesenchymal stem cells, MSCs) is considered to be one of the most promising seed cells for bone tissue engineering. It is speculated that FSS acting on hMSCs should produce biological effects similar to those of skeletal cells. In the experiment, the parallel plate flow chamber was modified to stimulate hMSCs with FSS, the morphological changes were observed by inverted microscope and transmission electron microscope, and cell proliferation was detected by MTT and flow cytometry. Osteogenic differentiation was determined by alkaline phosphatase and osteocalcin activity, alkaline phosphatase staining, calcium nodule alizarin red staining and tetracycline fluorescence labeling. By observing the effects of FSS on the proliferation and osteogenic differentiation of hMSCs in vitro, we provide a theoretical basis for further studying the mechanism of FSS stimulating proliferation and differentiation of hMSCs in vitro and using perfusion bioreactor to culture tissue engineered bone. The main findings and conclusions are as follows: First, the preparation and advantages of the improved parallel plate flow chamber. The flow chamber is filled with plexiglass, the plastic film is used to seal the flow chamber and keep the height of the flow chamber, the bottom of the plastic cell culture bottle is used as the cell attachment carrier, and the intravenous infusion tube is used as the perfusion fluid circulation pipe. The parallel plate flow chamber is fixed and sealed with a long tail clip. The whole device is sterilized by radiation irradiation. Compared with other reported fluid shear stimulation models, this model is simple to make, low cost and easy to operate. The experiment proves that the system is scientific and feasible as the research model of hMSCs hydrodynamics. Second, fluid shear stress can promote the proliferation of human bone marrow mesenchymal stem cells. Divide the third generation of hMSCs into
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R329.2

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 衛(wèi)元晨;基于微流控芯片對機(jī)械響應(yīng)細(xì)胞的研究[D];沈陽工業(yè)大學(xué);2013年

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本文編號：1930501