【摘要】：背景: 臨床上對(duì)于腫瘤等惡性疾病的治療手段,已從傳統(tǒng)的手術(shù)、放療、化療擴(kuò)展到了生物免疫治療。在這一領(lǐng)域中,細(xì)胞治療占主導(dǎo)地位。臍血以其豐富的干細(xì)胞、較弱的免疫細(xì)胞抗原性、簡單易行的采集和保存等多方面的優(yōu)勢,逐漸取代骨髓,而成為近年細(xì)胞治療中細(xì)胞的主要來源。因此臍血細(xì)胞的生物學(xué)性能、免疫學(xué)特性、殺傷腫瘤細(xì)胞(特別是血液系統(tǒng)腫瘤)的機(jī)制,成為了研究的熱點(diǎn)。 目的: 研究人臍血林巴細(xì)胞體外經(jīng)絲裂原刺激培養(yǎng)后的生物學(xué)、免疫學(xué)等方面的特性,并與健康人外周血淋巴細(xì)胞進(jìn)行比較,為臍血淋巴細(xì)胞輸注治療在臨床上的應(yīng)用提供客觀的指標(biāo)和理論基礎(chǔ)。 材料和方法: Ficoll密度梯度離心法分離的人臍血及外周血單個(gè)核細(xì)胞,體外無血清體系培養(yǎng)5天,同時(shí)設(shè)加PHA(植物血凝素)和不加PHA兩組。觀察細(xì)胞克隆生長情況,并在培養(yǎng)前和培養(yǎng)后不同時(shí)間點(diǎn)收集細(xì)胞和培養(yǎng)上清。臺(tái)盼蘭染色法計(jì)算活細(xì)胞數(shù);流式細(xì)胞術(shù)檢測細(xì)胞的表型特征;ELIsA法檢測培養(yǎng)上清中IFN-γ含量;Trizol提取細(xì)胞的RNA,逆轉(zhuǎn)錄成cDNA后,用FRQ-PCR法進(jìn)行IFN-γ及β-actin基因擴(kuò)增,相對(duì)定量地比較細(xì)胞內(nèi)IFN-γ mRNA轉(zhuǎn)錄以情況。 結(jié)果: 1.MNC體外生長情況:PHA(+)組UCBMNC在培養(yǎng)過程中細(xì)胞花團(tuán)的體積、數(shù)量、密集程度不斷增加;PBMNC細(xì)胞花團(tuán)的生長情況遠(yuǎn)不如UCBMNC旺盛。PHA(-)組UCBMNC在培養(yǎng)過程中未見明顯細(xì)胞花團(tuán),但細(xì)胞始終布滿培養(yǎng)孔;PBMNC無細(xì)胞花團(tuán),培養(yǎng)后期細(xì)胞分布甚至變得稀疏。 2.MNC細(xì)胞數(shù)量的擴(kuò)增:PHA(+)組UCBMNC細(xì)胞擴(kuò)增的最大值為
文內(nèi)圖片：
圖片說明：圖IUCBPHAG)組，，培養(yǎng)1天
[Abstract]:Background: the clinical treatment of malignant diseases such as tumor has been extended from traditional surgery, radiotherapy and chemotherapy to bioimmunotherapy. Cell therapy dominates in this field. Cord blood has gradually replaced bone marrow because of its rich stem cells, weak immune cell antigenicity, simple collection and preservation, and has become the main source of cells in cell therapy in recent years. Therefore, the biological performance and immunological characteristics of cord blood cells, and the mechanism of killing tumor cells (especially blood system tumors) have become the focus of research. Objective: to study the biological and immunological characteristics of human umbilical cord blood Limba cells stimulated by mitogen in vitro and to compare them with those of healthy peripheral blood lymphocytes, so as to provide an objective index and theoretical basis for the clinical application of cord blood lymphocytes infusion therapy. Materials and methods: human umbilical cord blood and peripheral blood mononuclear cells isolated by Ficoll density gradient centrifugation were cultured in serum-free system for 5 days. PHA (plant hemagglutinin) and no PHA were added at the same time. The cell clone growth was observed, and the cell and culture supernatant were collected at different time points before and after culture. The number of living cells was calculated by trypan blue staining, the phenotypic characteristics of the cells were detected by flow cytometry, the content of IFN- 緯 in the culture supernatant was detected by ELIsA assay, and the IFN- 緯 and 尾-actin genes were amplified by FRQ-PCR method after RNA, reverse transcription into cDNA, and the transcription of IFN- 緯 mRNA in the cells was compared relatively quantitatively. Results: the volume, number and density of UCBMNC cells in: PHA () group increased continuously during the culture process of 1.MNC in vitro, and the growth of PBMNC cells was much less exuberant than that of UCBMNC. PHA (-) group UCBMNC did not see obvious cell flower clusters during the culture process, but the cells were always covered with culture pores, and the cell distribution even became sparse in the later stage of culture. The maximum value of UCBMNC cell amplification in: PHA () group was as follows: the number of UCBMNC cells was amplified by the number of UCBMNC cells.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329.2

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