本文選題：組織工程 + B淋巴細(xì)胞��； 參考：《中國組織工程研究》2015年02期

【摘要】：背景:B淋巴細(xì)胞是機(jī)體免疫系統(tǒng)重要的參與者,目前的研究主要采用磁珠、補(bǔ)體等方法進(jìn)行B淋巴細(xì)胞的分離純化,但是這些方法費(fèi)用高或者細(xì)胞損傷大、純度低,體外分離、培養(yǎng)B淋巴細(xì)胞的方法還有待進(jìn)一步改進(jìn)。目的:探討從小鼠脾臟細(xì)胞中對B淋巴細(xì)胞同時(shí)進(jìn)行分離、培養(yǎng)的方法。采用加入白細(xì)胞介素4、脂多糖或者CD3單克隆抗體及其組合,探討體外分離、培養(yǎng)小鼠脾臟B淋巴細(xì)胞的適宜條件。方法:體外分離培養(yǎng)小鼠脾臟細(xì)胞,隨機(jī)分為7組,分別用白細(xì)胞介素4,CD3,脂多糖,白細(xì)胞介素4+CD3,白細(xì)胞介素4+脂多糖,CD3+脂多糖組進(jìn)行干預(yù)、將未給予刺激的脾細(xì)胞作為對照組。用流式細(xì)胞儀檢測在不同培養(yǎng)條件下小鼠脾臟細(xì)胞T、B淋巴細(xì)胞及其亞群的變化。結(jié)果與結(jié)論:與對照組比較,白細(xì)胞介素4組淋巴細(xì)胞在培養(yǎng)后第3-5天數(shù)量達(dá)到高峰;脂多糖組在培養(yǎng)初期無明顯作用,第3天開始淋巴細(xì)胞數(shù)量有明顯的增加,第5天達(dá)到高峰;培養(yǎng)體系中加入CD3單克隆抗體,可導(dǎo)致T淋巴細(xì)胞消失,培養(yǎng)2 d后,可得到較為單一的B淋巴細(xì)胞,其細(xì)胞數(shù)量在第3天達(dá)到高峰。其中B220+IgD+成熟B淋巴細(xì)胞亞群數(shù)量顯著增加。體外培養(yǎng)24 h后,各組B220+CD93+Transitional B淋巴細(xì)胞亞群均完全消失。結(jié)果說明,體外培養(yǎng)的小鼠脾臟細(xì)胞加入CD3單克隆抗體和白細(xì)胞介素4可以去除T淋巴細(xì)胞,并維持成熟B淋巴細(xì)胞的生存和增殖。
[Abstract]:Background: B lymphocytes are important participants in the body's immune system. Currently, the methods of separation and purification of B lymphocytes, such as magnetic beads and complements, are mainly used, but these methods are expensive or have high cell damage, low purity and in vitro isolation.The method of B lymphocyte culture needs further improvement.Objective: to study the method of simultaneous isolation and culture of B lymphocytes from mouse spleen cells.The suitable conditions for isolation and culture of mouse splenic B lymphocytes in vitro were studied by adding interleukin 4, lipopolysaccharide or CD3 monoclonal antibodies and their combinations.Methods: mouse spleen cells were isolated and cultured in vitro and randomly divided into 7 groups. The mice were treated with interleukin-4 CD3, lipopolysaccharide, interleukin-4 CD3, interleukin-4 lipopolysaccharide CD3 respectively.Splenocytes without stimulation were used as control group.The changes of T _ (B) lymphocytes and their subsets of spleen cells in different culture conditions were detected by flow cytometry.Results and conclusion: compared with the control group, the number of lymphocytes in the IL-4 group reached the peak on the 3-5 days after culture, but in the lipopolysaccharide group there was no obvious effect at the initial stage of culture, but the lymphocyte number increased significantly at the 3rd day.On the 5th day, the T lymphocytes disappeared when CD3 monoclonal antibody was added into the culture system. After 2 days of culture, a single B lymphocyte was obtained, and the number of B lymphocytes reached its peak on the third day.The number of mature B lymphocyte subsets of B 220 IgD increased significantly.After cultured in vitro for 24 h, B lymphocyte subsets of B 220 CD93 Transitional disappeared completely in each group.The results showed that murine spleen cells cultured in vitro with CD3 monoclonal antibody and interleukin 4 could remove T lymphocytes and maintain the survival and proliferation of mature B lymphocytes.
【作者單位】： 天津醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院生物化學(xué)與分子生物學(xué)系;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(81070271)~~
【分類號】：R392

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