本文關(guān)鍵詞： 臍帶 間充質(zhì)干細胞 組織工程 接種方法 循環(huán)灌注　出處：《蘭州大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：人臍帶間充質(zhì)干細胞在骨組織工程領(lǐng)域有廣泛的應(yīng)用前景。三維載體上細胞的接種方法對于骨組織構(gòu)建有重要的影響。本文應(yīng)用自制的循環(huán)灌注接種系統(tǒng)研究循環(huán)灌注接種,以及離心接種和靜態(tài)接種這三種接種方法在人臍帶間充質(zhì)干細胞骨組織工程領(lǐng)域的應(yīng)用。 培養(yǎng)傳代人臍帶間充質(zhì)干細胞,流式細胞儀檢測細胞表面標記,將細胞用循環(huán)灌注方法,離心接種方法和靜態(tài)接種方法接種至三維聚對苯二甲酸乙二醇酯(Polyethylene terephthalate, PET)無紡布片狀載體,CCK-8細胞計數(shù)法計算接種效率,乳酸脫氫酶法測定細胞損傷,連續(xù)培養(yǎng)18天并繪制生長曲線。18天后使用成骨誘導(dǎo)分化培養(yǎng)液連續(xù)誘導(dǎo)分化20天,檢測堿性磷酸酶活性比較不同接種方法的細胞成骨分化能力。結(jié)果顯示傳代細胞形態(tài)穩(wěn)定、活力好,高表達CD90,CD105,不表達CD14,CD45。循環(huán)灌注方法組和離心接種方法組的接種效率,細胞密度,增殖能力均優(yōu)于靜態(tài)接種組。與離心接種方法和靜態(tài)接種方法相比循環(huán)灌注組LDH偏高,并且細胞的延遲期長于靜態(tài)接種。循環(huán)灌注方法組和離心方法組的堿性磷酸酶活性高于靜態(tài)接種組。結(jié)果表明,循環(huán)灌注接種和離心接種方法組更適合間充質(zhì)干細胞骨組織工程的應(yīng)用,但進一步應(yīng)用仍需對工程參數(shù)進行優(yōu)化。
[Abstract]:Human umbilical cord mesenchymal stem cells have a wide application prospect in bone tissue engineering. The method of cell inoculation on three-dimensional carrier has an important effect on bone tissue construction. Ring perfusion inoculation. And the application of centrifugation and static inoculation in bone tissue engineering of human umbilical cord mesenchymal stem cells. Human umbilical cord mesenchymal stem cells were cultured and the surface markers were detected by flow cytometry. Centrifugal and static inoculation methods were used to inoculate the three-dimensional polyethylene terephthalate (PET) non-woven sheet carrier. CCK-8 cell count method was used to calculate the inoculation efficiency, and lactate dehydrogenase method was used to determine the cell damage. After 18 days of continuous culture, the growth curve was drawn and the osteogenic differentiation medium was used to induce differentiation for 20 days. The activity of alkaline phosphatase was compared with the osteogenic differentiation ability of different inoculation methods. The results showed that the passaged cells were stable in morphology and good in activity. CD90 CD105 was highly expressed and CD14 was not expressed. CD45. the inoculation efficiency and cell density of circulatory perfusion group and centrifugal inoculation group. The proliferative ability was superior to the static inoculation group, and the LDH of the circulatory perfusion group was higher than that of the centrifugal inoculation method and the static inoculation method. The activity of alkaline phosphatase in the circulatory perfusion group and centrifugation group was higher than that in the static inoculation group. The methods of circulatory perfusion inoculation and centrifugal inoculation were more suitable for the application of bone tissue engineering of mesenchymal stem cells, but further application still needed to optimize the engineering parameters.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329

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本文編號：1493697