Apoptosis Induction and Detection in a Primary Culture of Sea Cucumber Intestinal Cells.
Apoptosis Induction and Detection in a Primary Culture of Sea Cucumber Intestinal Cells.
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DOI：
10.3791/60557
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发表时间：
2020-01
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影响因子：
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通讯作者：
Tianming Wang;Xu Chen;Ke Xu;Bing Zhang;Dexiang Huang;Jingwen Yang
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文献类型：
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作者：
Tianming Wang;Xu Chen;Ke Xu;Bing Zhang;Dexiang Huang;Jingwen Yang
原代培养细胞在各种科学学科中被用作生物物质功能评估或特定生物活动表征的特别重要的工具。然而，由于缺乏普遍适用的细胞培养介质和方案，对海洋生物进行细胞培养的方法仍然有限。同时，海洋无脊椎动物细胞普遍存在的微生物污染和多变性进一步阻碍了建立有效的海洋无脊椎动物细胞培养策略。在这里，我们描述了一种简便的刺参肠道细胞培养方法；此外，我们还提供了一个体外诱导和检测原代培养的肠道细胞凋亡的例子。此外，本实验还提供了合适的培养基和细胞采集方法的详细信息。所描述的方案与从棘皮动物、软体动物和甲壳动物等海洋生物中广泛获得的各种组织样本相兼容，并且它可以为多种体外实验应用提供足够的细胞。这项技术将使研究人员能够有效地操纵海洋无脊椎动物的原代细胞培养，并有助于对细胞上的目标生物材料进行功能评估。
Primary cultured cells are used in a variety of scientific disciplines as exceptionally important tools for the functional evaluation of biological substances or characterization of specific biological activities. However, due to the lack of universally applicable cell culture media and protocols, well described cell culture methods for marine organisms are still limited. Meanwhile, the commonly occurring microbial contamination and polytropic properties of marine invertebrate cells further impede the establishment of an effective cell culture strategy for marine invertebrates. Here, we describe an easy-to-handle method for culturing intestinal cells from sea cucumber Apostichopus japonicus; additionally, we provide an example of in vitro apoptosis induction and detection in primary cultured intestinal cells. Moreover, this experiment provides details about the appropriate culture medium and cell collection method. The described protocol is compatible with a variety of widely available tissue samples from marine organisms including Echinodermata, Mollusca, and Crustacea, and it can provide sufficient cells for multiple in vitro experimental applications. This technique would enable researchers to efficiently manipulate primary cell cultures from marine invertebrates and to facilitate the functional evaluation of targeted biological materials on cells.