Epithelial cell cultures from Botryllus schlosseri palleal buds: accomplishments and challenges.
Epithelial cell cultures from Botryllus schlosseri palleal buds: accomplishments and challenges.
复制标题
灰霉病菌苍白芽的上皮细胞培养：成就和挑战。
DOI：
10.1007/s11022-004-2087-9
复制
发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Rinkevich,Baruch
中科院分区：
文献类型：
--
作者：
Rabinowitz,Claudette;Rinkevich,Baruch
本研究的重点是最近的改进，上皮单层培养源自整个提取的botryllus schlosseri（Urochordata）芽。芽（n= 2000）在不同的（‘ A ’到‘ D ’）胚发育阶段取。我们测试了35种不同基质和培养基组合在附着、细胞扩散、上皮生长频率和单层寿命方面的适用性。在良好的条件下，胚性阶段B到D（而不是A阶段）的培养芽在3天的过渡期后开始附着在底物上，形成球体和附着的单层。衬底类型对单分子膜的附着和发展至关重要。在不同的培养条件下，‘B’期和‘C’期的一些芽会在3-20天内形成一个或多个直径为1毫米的大球体。阶段‘ D ’芽发育单层（高达芽的20%），不经过球体阶段。在A期芽培养中未观察到球形上皮和附着单层上皮。使用特定的培养基类型，球体以每天60 μm直径的速度生长，除非存在合适的底物，否则不会附着。粘附后，上皮单层以每天200 μm直径的速度扩张，持续3-15天，随后分离并死亡。共测试了16种介质。发现培养基和底物组合决定上皮细胞的寿命。这些结果表明，从葡萄芽中培养上皮单层有显著的改善。然而，发育的上皮层的早期衰老（从出现开始长达两周）可能表明内部老化时钟，应该在未来的方法中考虑。
This study focuses on recent improvement in epithelial monolayer cultures originating from whole extirpatedBotryllus schlosseri(Urochordata) buds. Buds (n= 2,000) were taken at different (‘A’ to ‘D’) blastogenic stages. We tested the suitability of 35 combinations of various substrates and media on attachment, cell spread, epithelial growth frequencies and on monolayer lifespans. Under favorable conditions, cultured buds at blastogenic stages ‘B’ to ‘D’ (but not stage ‘A’) started to attach to the substrates following a 3-day transient period that leads to formation of spheres and attached monolayers. Substrate type is important for the attachment and the development of monolayers. Under various culture conditions, some of stages ‘B’ and ‘C’ buds develop (3–20 days) one or more large (1 mm diameter) spheres. Stage ‘D’ buds develop monolayers (up to 20% of buds) without going through a sphere phase. Neither spheres nor attached monolayers of epithelium were observed in stage ‘A’ bud cultures. Spheres grew at a rate of 60 μm in diameter per day using specific medium types and did not attach unless the appropriate substrate was present. When attached, epithelial monolayers expanded at a rate of 200 μm in diameter per day, for 3–15 days, and subsequently detached and died. Sixteen types of media were tested. Medium and substrate combinations were found to determine epithelial lifespan. These results revealed significant improvements in the culture of epithelial monolayers fromBotrylluspalleal buds. However, an early senescence of the developed epithelial sheets (up to two weeks from onset of appearance) may indicate an internal ageing clock that should be taken into consideration in future approaches.