Induction of ciliated cells from avian embryonic stem cells using three-dimensional matrix.
Induction of ciliated cells from avian embryonic stem cells using three-dimensional matrix.
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DOI：
10.1089/ten.tec.2009.0327
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发表时间：
2010-02
期刊：
影响因子：
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通讯作者：
Yuchi Wang;L. Wong;H. Mao
中科院分区：
文献类型：
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作者：
Yuchi Wang;L. Wong;H. Mao
我们设计了一种简单的三维（3D）组织培养方法，通过使用鸟类受精卵诱导鸟类胚胎干（ES）细胞的纤毛发生。与以前报道的技术不同，该方法不需要胰蛋白酶消化，这会降低细胞的活力;它也不需要气液界面来诱导纤毛发生和维持诱导的纤毛细胞的生长。ES细胞接种并附着在这种胶原包被的壳聚糖3D凝胶上自发地生长，并且生长健壮。在抑制胚状体形成的情况下培养2周后，观察到具有明显且有力的纤毛跳动的细胞。我们测量了这些ES分化的纤毛细胞的纤毛搏动频率40天。这些结果与我们以前研究中对其他物种（包括人类）纤毛细胞的所有报告测量结果一致。这些数据意味着，这些ES衍生的纤毛细胞，在其固有的基础自主节律率跳动的纤毛，保持纤毛节拍频率的监管机制的完整性。总之，我们已经表明，在3D组织工程支架中培养的ES细胞是一种很有前途的方法，用于开发密切模仿体内纤毛细胞自然环境的体外细胞模型。这种细胞模型可能是基于细胞的高通量筛选和发现肺部药物的纤毛细胞的来源。
We have devised a simple three-dimensional (3D) tissue-culturing method to induce ciliogenesis from avian embryonic stem (ES) cells by using avian fertilized eggs. Unlike the previous reported techniques, this method does not require trypsinization, which would reduce the viability of the cells; it also does not require an air-liquid interface to induce ciliogenesis and to maintain the growth of the induced ciliated cells. ES cells seeded and attached on this collagen-coated chitosan 3D gel grew spontaneously and robustly. Following 2 weeks in culture with inhibition of embryoid body formation, cells with noticeable and vigorous beating cilia were observed. We measured the ciliary beat frequencies of these ES-differentiated ciliated cells for 40 days. These results were consistent with all reported measurements made for other species of ciliated cells, including human, from our previous study. These data imply that the cilia of these ES-derived ciliated cells, beating at their intrinsic basal autorhythmic rate, preserve the integrity of the regulatory mechanisms of ciliary beat frequency. In conclusion, we have shown that ES cells cultured in a 3D tissue-engineered scaffold is a promising approach for developing an in vitro cell model that closely mimics the in vivo ciliated cell natural milieu. This cell model can potentially be the source of ciliated cells for cell-based high-throughput screening and discovery of pulmonary drugs.