Formation of Human Fibroblast Aggregates (Spheroids) by Rotational Culture
Formation of Human Fibroblast Aggregates (Spheroids) by Rotational Culture
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通过旋转培养形成人成纤维细胞聚集体（球体）
DOI：
10.3727/000000001783986503
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发表时间：
2001
影响因子：
3.3
通讯作者：
T. Tateishi
中科院分区：
文献类型：
--
作者：
K. Furukawa;T. Ushida;Y. Sakai;Motoyuki Suzuki;J. Tanaka;T. Tateishi
在目前的研究中，我们试图通过旋转摇动，减少成纤维细胞-材料相互作用，并增加细胞-细胞相互作用来形成成纤维细胞聚集体。此外，为了促进细胞-细胞相互作用，培养基补充有胰岛素、地塞米松和碱性成纤维细胞生长。在这种改进的培养条件下，正常新生人皮肤成纤维细胞在旋转摇床上旋转1天内形成球状聚集体。形成的聚集体具有不规则的形状，并且在12小时后仅由几个细胞组成。然而，在摇动24小时后，它们变成近球形。聚集体直径约为240 μm。经36 h振荡后，其形状变得更圆，表面变得更光滑。在聚集体的中心没有观察到坏死的证据，尽管少量的死细胞散布在整个聚集体中。24-36小时后，收集正常人成纤维细胞的聚集体，并重新接种到由聚乙醇酸组成的支架上，聚乙醇酸在商业上用作涂覆有胶原的人造皮肤的支架。聚集体被成功地捕获到聚乙醇酸的网，并在24小时内附着。因此，聚集体可以提供用于将成纤维细胞接种到人造皮肤支架（例如聚乙醇酸网）的替代方法。
In the current study, we attempted to form aggregates of fibroblasts by rotationally shaking, declining fibroblast–material interactions, and augmenting cell–cell interactions. In addition, to promote cell–cell interactions, the medium was supplemented with insulin, dexamethasone, and basic fibroblast growth. Under such improved culture conditions, normal neonatal human dermal fibroblasts formed spheroidal aggregates within 1 day of rotation on a rotational shaker. The aggregates that formed had irregular shapes and were composed from only several cells after 12 h. However, they became nearly spheroidal after 24 h of shaking. The aggregates were approximately 240 μm in diameter. After 36 h of shaking, their shape became more rounded and their surfaces became smoother. No evidence of necrosis in the center of the aggregates was observed, although a small number of dead cells was scattered throughout the aggregates. After 24–36 h, aggregates of normal human fibroblasts were collected and reinoculated onto a scaffold composed of polyglycolic acid, which is used commercially as a scaffold for artificial skin, coated with collagen. The aggregates were successfully trapped to the mesh of polyglycolic acid and became attached within 24 h. Therefore, the aggregates could provide an alternative method for seeding fibroblasts to scaffold for an artificial skin, such as a mesh of polyglycolic acid.