Giordano Gennaro, Hong Sungwoo, Faustman Elaine M, Costa Lucio G
Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2011;758:171-8. doi: 10.1007/978-1-61779-170-3_11.
During brain development, cell death is a physiological process which allows the elimination of cells produced in excess. During adulthood, when there is no or little physiologic cell death, an increase in cell loss is usually caused by neurologic disorders or by exposure to neurotoxic chemicals. Measurements of cell death are often used a first line of investigation on chemicals. Cell death in neuronal or glial cultures in vitro can be quantified with a variety of assays based on different properties of live and dead cells. Thus, healthy cells exclude dye (e.g., trypan blue, propidium iodide) or possess metabolic activity to cause a compound's conversion to a colored or fluorescent one (e.g., MTT, calcein AM) while dead cells do not. Conversely, dying cells release enzymes in the medium (e.g., LDH) whose quantification is proportional to the number of dead cells. This chapter describes several relatively rapid, inexpensive and reliable methods for measuring cell death and in neurons and astrocytes in primary cultures or in neuronal and glial cell lines.
在大脑发育过程中，细胞死亡是一个生理过程，它能使过量产生的细胞被清除。在成年期，当没有或仅有少量生理性细胞死亡时，细胞损失的增加通常是由神经疾病或接触神经毒性化学物质引起的。细胞死亡的检测常常被用作对化学物质进行初步调查的手段。体外神经元或神经胶质细胞培养中的细胞死亡可以通过基于活细胞和死细胞不同特性的多种检测方法进行量化。因此，健康细胞会排斥染料（如台盼蓝、碘化丙啶）或具有代谢活性，能使化合物转化为有颜色或荧光的物质（如MTT、钙黄绿素AM），而死细胞则不会。相反，濒死细胞会向培养基中释放酶（如乳酸脱氢酶），其定量与死细胞数量成正比。本章介绍了几种相对快速、廉价且可靠的方法，用于测量原代培养物或神经元及神经胶质细胞系中神经元和星形胶质细胞的细胞死亡情况。
