Heckman C A
In Vitro. 1983 Jan;19(1):31-40. doi: 10.1007/BF02617991.
Previous work in this laboratory demonstrated that rat respiratory airway epithelial cells grown from tissue explants undergo concurrent division and differentiation in culture. In the present studies, this model system has been used to optimize conditions for epithelial growth, with the goal of facilitating the culture of epithelium from other organs and species. Fibroblasts appeared to limit the expansion of epithelial outgrowths; their growth was controlled by using delipidated serum, rather than serum, in the culture media, with addition of adrenergic antagonists. In nonsupplemented media, rat tracheal fibroblasts doubled in number in about 3 d. The doubling time was slowed to more than 9 d by the serum substitution in conjunction with the alpha-antagonist phenoxybenzamine. Rat tracheal epithelial cultures maintained under identical conditions doubled in less than 4 d, so that a growth advantage of threefold was achieved. The combination of conditions also seemed to inhibit growth of fibroblasts from other species. However, even when the problem of fibroblast overgrowth was obviated, the respiratory airway epithelia of the mouse, hamster, and marmoset failed to show self-sustaining growth in culture. The rat epithelium continued to grow for over 6 wk. A number of other organs from the rat, notably esophagus, skin, and salivary gland, showed self-sustaining growth after removal of explants. Although epithelial outgrowths were formed by explants from certain organs of the hamster, only those from the kidney and thyroid continued to grow after removal of the tissue explants. Therefore, growth failure in cultured epithelia, quantitatively defined, is common and frequently unrelated to the problem of fibroblast overgrowth. The rat seems to be the species of choice for studies on the isolated epithelia from most organ sites.
本实验室先前的研究表明,从组织外植体生长而来的大鼠呼吸道上皮细胞在培养过程中会同时进行分裂和分化。在本研究中,该模型系统已被用于优化上皮细胞生长条件,目的是促进其他器官和物种上皮细胞的培养。成纤维细胞似乎限制了上皮细胞生长物的扩展;通过在培养基中使用脱脂血清而非血清,并添加肾上腺素能拮抗剂来控制其生长。在未添加血清的培养基中,大鼠气管成纤维细胞数量在约3天内翻倍。通过血清替代结合α-拮抗剂苯氧苄胺,细胞翻倍时间延长至超过9天。在相同条件下培养的大鼠气管上皮细胞数量在不到4天内翻倍,从而实现了三倍的生长优势。这些条件的组合似乎也抑制了其他物种成纤维细胞的生长。然而,即使解决了成纤维细胞过度生长的问题,小鼠、仓鼠和狨猴的呼吸道上皮细胞在培养中仍未能显示出自给自足的生长。大鼠上皮细胞持续生长超过6周。大鼠的许多其他器官,特别是食管、皮肤和唾液腺,在去除外植体后显示出自给自足的生长。虽然仓鼠某些器官的外植体形成了上皮细胞生长物,但只有肾脏和甲状腺的外植体在去除组织外植体后继续生长。因此,定量定义的培养上皮细胞生长失败很常见,且通常与成纤维细胞过度生长问题无关。对于大多数器官部位分离上皮细胞的研究,大鼠似乎是首选物种。