菇蕈多醣體對體外培養之南美白蝦血淋巴細胞之影響

The effect of β-glucan on the in vitro haemocytes of Litopenaeus vannamei

梁栩福(Eddie Shu Pu, Leong)

指導教授 ： 陳秀男

國立臺灣大學/生命科學院/漁業科學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.01326

摘要

本研究主要探討菇蕈多醣體（mushroom β-glucan）對體外培養之南美白蝦血淋巴細胞之影響。本實驗將南美白蝦血淋巴細胞進行初代培養（primary culture），結果發現調整滲透壓之培養基組別顯著提高了南美白蝦血淋巴細胞活性並延長存活時間。培養基添加多醣體之後發現，0.5%濃度組別可以延長血淋巴細胞活性維持。將滲透壓調整為900±5 mOsm/Kg 的培養基中添加多醣體，發現可以降低南美白蝦血淋巴細胞凋亡比例；細胞中的ROS產生率會隨著多醣體濃度提升，會有下降再上升的情形；細胞吞噬作用有顯著性的提升，且細胞大小與顆粒性會有複雜化的現象。結果也發現，血淋巴細胞於體外培養時會有類似黑化作用及胞封作用的情形發生，並且隨著多醣體刺激濃度的升高，三種細胞的顆粒性與大小以流式細胞儀監測觀察下有趨向同一種型態的趨勢。

關鍵字

南美白蝦 ； 血淋巴細胞 ； 細胞培養 ； 滲透壓 ； 流式細胞儀 ； 菇蕈多醣體

並列摘要

The present study aims to examine the effects of mushroom β-glucan on the cultured Litopenaeus vannamei haemocytes via in-vitro experiment. The cell viability and activity were enhanced, while the osmolarity of cell cultured medium was adjusted into the 900±5mOsm/Kg compared to the unadjusted （315±5mOsm/Kg） cultured medium. Further examination also presented that the adjusted cell cultured medium additional supplemented with the 0.5% β-glucan induce the prolonger of the cell survival compared to the unadjusted group. In the ROS production and apoptosis examination, while the shrimp haemocytes cultured in the adjusted cell cultured medium supplemented with the β-glucan presented that apoptosis cituation of the haemocyte was to be reduced but the ROS production was to be enhanced. The phagocytosis ability, cell particles and sizes of the haemocyte cultured in the adjusted cell culture medium supplemented with β-glucan, we observed that the complication of the intracellular particle and the cell size were to be complicated via the FLOW Cytometry observation and the phagocytosis ablity was to be increased.
In this research, we observed that the osmolarity of the shrimp cultured medium was supposed to be adjusted in to 900 ±5mOsm/Kg to maintain the cell viability and activity compared to the unadjusted culture medium in the lab experiment. And we also presented that the β-glucan exactly induce the survival rate, phagocytosis activity and the ROS production of shrimp haemocytes, but the reduce the apoptosis of the shrimp haemocytes via the in-vitro experiment.

並列關鍵字

white shrimp ； haemocytes ； cell culture ； osmolarity ； flow cytometric ； β-glucan

參考文獻

吳協峻，2006。葡聚多醣體與藻酸鈉對斷溝龍蝦非特異性免疫反應的影響。國立

龔子棋，2011。小柴胡湯對南美白蝦非特異性免疫反應之影響。國立臺灣海洋大

Lin, M. N., Ting, Y. Y., Tzeng, B. S., Liu, C. Y. 1990. Penaeid parental shrimp rearing: culture of the third generation in Penaeus vannamei. Journal of Fisheries Society of Taiwan. 17, 125-132.

王婉媜，2013。菇蕈多醣體對於原發位腫瘤微環境之影響。國立台灣大學漁業科

吳育昇，2014。活性氧物質對吳郭魚肝臟駐留巨噬細胞與組織修復。國立台灣大