Guerrero R R, Rounds D E, Booher J
In Vitro. 1977 Aug;13(8):517-24. doi: 10.1007/BF02615145.
An improved method for maintaining adult rat lung in submerged organ culture is described in which the alveoli were inflated with agar and 200-micron-thick hand-cut sections were mounted in Rose chambers. The conventional single-compartmented Rose culture chamber was modified by adding a second chamber separated from the first by a gaspermeable membrane. One compartment functioned as an air reservoir and the other housed the explants submerged in nutrient medium. Visking dialysis membrane used underneath the explants prevented cell outgrowth and facilitated the exchange of nutrients and waste products at the glass-tissue interface. Because of the excellent optical properties of the Rose chamber and the thinness of the explants, individual cell types can be identified in the living tissue. The explants were studied with time-lapse cinematography, light microcoscopy, histology, and erythrosine B for dye exclusion. With this modified system the functional life span of the explants was increased from 1 week to 1 month.
本文描述了一种改进的成年大鼠肺器官浸没培养方法，其中肺泡用琼脂充气，将200微米厚的手工切片置于罗斯培养室中。传统的单隔层罗斯培养室经过改进，增加了一个由透气膜与第一个隔层隔开的第二个隔层。一个隔层用作空气储存器，另一个隔层容纳浸没在营养培养基中的外植体。外植体下方使用的维斯金透析膜可防止细胞生长，并促进玻璃-组织界面处营养物质和代谢产物的交换。由于罗斯培养室具有出色的光学特性且外植体很薄，因此可以在活组织中识别出不同的细胞类型。通过延时摄影、光学显微镜、组织学和赤藓红B染料排除法对外植体进行了研究。使用这种改进系统，外植体的功能寿命从1周延长到了1个月。
