In vitro culture of blood cells from the colonial protochordateBotryllus schlosseri
In vitro culture of blood cells from the colonial protochordateBotryllus schlosseri
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群体原索动物血细胞的体外培养
DOI：
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
C. Rabinowitz
中科院分区：
文献类型：
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作者：
B. Rinkevich;C. Rabinowitz
本文报道了在96孔板中培养施氏葡萄球菌（Botryllusschlosseri）循环血细胞3个月的方法。在基于Dulbecco改良的Eagle培养基的培养基中，补充有葡萄状海鞘血淋巴渗透压的盐、HEPES缓冲液、l-维生素A、胎牛血清和抗生素。颗粒色素细胞间的细胞桥在24 h内建立。这些细胞的活力下降缓慢，大多数在1个月内死亡。没有任何细胞增殖的迹象其他细胞类型保持在停滞状态，并进行每周培养基更换。从2 wk到1 mo，在6 - 10%的威尔斯孔中随机记录自发细胞增殖。这种增殖之后形成大量细胞团块，从中均匀的血细胞（5 µm，淋巴细胞样细胞）向外周迁移。应激条件，包括培养基更换和部分培养基更换之间的较长间隔，增加了细胞增殖的可能性。从每个增殖的原代培养物，我们成功地进行了10个铺板周期超过15周的时间，在此期间，细胞分化的大小，但均匀的结构。这产生了第一个葡萄状淋巴细胞样细胞系。从这个阶段开始，细胞数量保持恒定长达6个月。而不增加细胞数量。几种促有丝分裂因子的原代culture.Botryllus和海参血淋巴细胞和混合白细胞介素被发现增加显着增殖的至少一个特定的细胞大小，而细胞没有显着响应凝集素（Concavalin A，麦胚凝集素，荆豆凝集素），胰岛素和视黄酸。结果进行了讨论与未来的努力，在被囊血细胞培养的发展。
SummaryPrimary cultures of circulatory blood cells from the colonial tunicateBotryllus schlosseri were cultivated in 96-well plates for up to 3 mo. in a medium based on Dulbecco’s modified Eagle’s medium, supplemented with salts to the botryllid ascidian hemolymph osmolarity, HEPES buffer,l-glutamin, fetal bovine serum, and antibiotics. Intercellular bridges between granular pigment cells were established within 24 h. The viability of these cells decreased slowly, and most died within 1 mo. without any sign of cell proliferation. Other cell types remained in an arrested state and were subjected to a weekly medium exchange. Spontaneous cell proliferation was randomly recorded in 6 to 10% of the wells from 2 wk to 1 mo. This proliferation was followed by the formation of masses of cell clumps, from which uniform hemocytes (5 µm, lymphocytelike cells) migrated peripherally. Stress conditions, which included longer intervals between medium exchanges and partial medium replacement, increased the probability of cell proliferation. From each proliferating primary culture, we successfully performed up to 10 plating cycles over a period of 15 wk, during which the cells differentiate in size but are uniformly structured. This produced the firstBotryllus lymphocytelike cell line. From this stage, cell numbers remained constant for up to 6 mo. without increase in cell number. Several mitogenic factors were employed on primary cultures.Botryllus and sea cucumber hemolymphs and mixed interleukins were found to augment significantly proliferation of at least one specific cell size, whereas cells were not markedly responsive to lectins (Concavalin A, wheat germ agglutinin,Ulex europaeus agglutinin), insulin, and retinoic acid. The results are discussed with respect to future efforts in the development of tunicate blood cell cultures.