O'Malley M B, MacLeish P R
Rockefeller University, New York, NY 10021.
J Neurosci Methods. 1993 Apr;47(1-2):61-71. doi: 10.1016/0165-0270(93)90022-j.
A culture system for adult monkey and human retinal cells was developed utilizing monoclonal antibodies as substrates. Monoclonal antibodies were made using monkey retinal membranes as immunogen, and screening for the ability to bind and facilitate growth of dissociated retinal neurons when used as a culture substrate. Two antibodies, MR1 and 9B5, which recognize immunohistochemically distinct cell surface antigens, were found to work effectively as a culture substrate for dissociated monkey retinal cells. A variety of dissociated monkey retinal neurons and glial cells grew extensive processes on MR1 and 9B5, and continued to express cell-specific antigens. In addition, many multipolar neurons remained electrically excitable after several days in culture. Dissociated human retinal cells did not cross-react with MR1 but grew well on 9B5. These results extend previous work showing that antibody substrates can support the growth of neurons derived from other vertebrate species (e.g., neonatal rat, adult salamander). The long-term survival and growth of primate cells in culture supports the current view that neurons in the adult mammalian central nervous system (CNS) may regenerate under permissive circumstances. Further, these data provide evidence that direct support by non-neuronal cells is not required and that regeneration in vitro may be mediated by suitably adhesive non-cellular substrates.
利用单克隆抗体作为底物开发了一种用于成年猴和人类视网膜细胞的培养系统。以猴视网膜膜作为免疫原制备单克隆抗体,并筛选其用作培养底物时结合和解离视网膜神经元并促进其生长的能力。发现两种抗体MR1和9B5可有效用作解离的猴视网膜细胞的培养底物,它们在免疫组织化学上识别不同的细胞表面抗原。多种解离的猴视网膜神经元和神经胶质细胞在MR1和9B5上长出了广泛的突起,并继续表达细胞特异性抗原。此外,许多多极神经元在培养几天后仍保持电兴奋性。解离的人类视网膜细胞与MR1无交叉反应,但在9B5上生长良好。这些结果扩展了先前的研究工作,表明抗体底物可以支持源自其他脊椎动物物种(如新生大鼠、成年蝾螈)的神经元的生长。灵长类细胞在培养中的长期存活和生长支持了当前的观点,即成年哺乳动物中枢神经系统(CNS)中的神经元在允许的情况下可能会再生。此外,这些数据提供了证据,表明不需要非神经元细胞的直接支持,并且体外再生可能由合适的粘附性非细胞底物介导。