Darmon M Y
In Vitro. 1982 Dec;18(12):997-1003. doi: 10.1007/BF02796374.
Culture of cells in hormonally defined media has allowed (a) the demonstration of physiological responses from cells usually unable to express them in vitro and (b) the study of the effects on growth and differentiation of diffusible factors and attachment factors. The embryonal carcinoma line 1003 forms multidifferentiated tumors in vivo but is unable to differentiate in vitro when grown in serum-containing medium. In a defined medium containing insulin, transferrin, selenium, and fibronectin as attachment factors, 1003 cells grow for several generations and differentiate into neurons and embryonic mesenchyme (Darmon et al., 1981, Dev. Biol. 85: 463-473). In the present work the effects of fibronectin and laminin were compared. In the presence of laminin the cells attached and spread better, grew faster, and could be plated at lower densities. Neurite extension was also better under these conditions and most importantly, it was found that laminin induced an important formation of muscular tissue when the cells had been seeded at low densities. Multinucleated myotubes could be stained with antibodies directed against embryonic muscular myosin. Coating the dishes with polylysine or adding FGF or serum-spreading factor to the medium allowed growth of low-density cultures with fibronectin instead of laminin but muscular differentiation was not detected under these conditions. Addition of fibronectin to laminin-containing medium did not inhibit muscular differentiation.
在激素限定培养基中培养细胞，使得（a）通常在体外无法表达生理反应的细胞能够展现出这些反应，以及（b）能够研究可扩散因子和黏附因子对生长和分化的影响。胚胎癌细胞系1003在体内形成多分化肿瘤，但在含血清培养基中培养时在体外无法分化。在含有胰岛素、转铁蛋白、硒和纤连蛋白作为黏附因子的限定培养基中，1003细胞能生长几代并分化为神经元和胚胎间充质（达蒙等人，1981年，《发育生物学》85卷：463 - 473页）。在本研究中，比较了纤连蛋白和层粘连蛋白的作用。在层粘连蛋白存在的情况下，细胞附着和铺展得更好，生长更快，并且可以以更低的密度接种。在这些条件下神经突延伸也更好，最重要的是，发现当细胞以低密度接种时，层粘连蛋白诱导了肌肉组织的重要形成。多核肌管可用针对胚胎肌肉肌球蛋白的抗体进行染色。用聚赖氨酸包被培养皿或向培养基中添加成纤维细胞生长因子或血清扩散因子，可使含纤连蛋白而非层粘连蛋白的低密度培养物生长，但在这些条件下未检测到肌肉分化。向含层粘连蛋白的培养基中添加纤连蛋白并不抑制肌肉分化。
