Hosokawa Takeshi, Betsuyaku Tomoko, Nishimura Masaharu, Furuyama Akiko, Katagiri Kazuko, Mochitate Katsumi
First Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
Connect Tissue Res. 2007;48(1):9-18. doi: 10.1080/03008200601017488.
Although lung epithelial cells directly attach to the basement membrane underneath in vivo, harvested epithelial cells are typically cultured on type I collagen gel (Col I-gel) in vitro. Recently we developed new culture substratum, designated as "synthesized Basement Membrane" (sBM), that has bared lamina densa on fibrillar collagen. To validate the usefulness of sBM substratum in airway tissue reconstitution in vitro, we cultured rat tracheal epithelial cells on sBM substratum and Col I-gel. When starting the air-liquid interface culture, most of the epithelial cells were squamous and positive for the basal cell marker cytokeratin 14 (CK14). After 14 days on sBM substratum, CK14-positive cells differentiated not only to Clara and mucous cells, but also to ciliated cells. Those differentiated cells formed pseudostratified-like epithelium and the remaining CK14-positive cells were polarized to the basal side. However, on Col I-gel, the CK14-positive cells were still squamous and not polarized, and ciliated cells did not appear. In conclusion, we established a new culture model on sBM substratum in which basal cells could differentiate to ciliated cells. The application of sBM substratum is useful in the study of the airway epithelial cell differentiation in vitro.
虽然肺上皮细胞在体内直接附着于其下方的基底膜，但收获的上皮细胞通常在体外培养于I型胶原凝胶（Col I - gel）上。最近我们开发了一种新的培养基质，命名为“合成基底膜”（sBM），其在纤维状胶原上具有裸露的致密层。为了验证sBM基质在体外气道组织重建中的实用性，我们将大鼠气管上皮细胞培养在sBM基质和Col I - gel上。开始气液界面培养时，大多数上皮细胞呈鳞状，基底细胞标志物细胞角蛋白14（CK14）呈阳性。在sBM基质上培养14天后，CK14阳性细胞不仅分化为克拉拉细胞和黏液细胞，还分化为纤毛细胞。这些分化的细胞形成假复层样上皮，其余CK14阳性细胞极化至基底侧。然而，在Col I - gel上，CK14阳性细胞仍为鳞状且未极化，纤毛细胞未出现。总之，我们在sBM基质上建立了一种新的培养模型，其中基底细胞可分化为纤毛细胞。sBM基质的应用在体外气道上皮细胞分化研究中是有用的。
