Baur H, Kasperek S, Pfaff E
Hoppe Seylers Z Physiol Chem. 1975 Jun;356(6):827-38. doi: 10.1515/bchm2.1975.356.s1.827.
36.4 +/Various cellular parameters were measured with regard to their usefulness as criteria of viability of isolated cells. Stainability by trypan blue and release of lactate dehydrogenase indicate only severe irreversible damage of cells. Neither endogenous respiration nor even the ATP/ADP ratio is a sensitive criterion of viability. On aging of cells, the ATP/ADP ratio remains high, even though the membrane potential, the intracellular K concentration and the content of adenine nucleotides decrease considerably. A sensitive, easily performed test is the stimulation of cellular respiration by 1mM succinate. Only a damaged plasma membrane allows succinate permeation of a rate sufficient to stimulate respiration. The membrane potential and the intracellular Na and K concentrations are the most sensitive criteria of viability, since they indicate the earliest changes on aging. (For freshly isolated cells, we found a membrane potential of 36.4 "/- 3.4 mv [n = 5], an intracellular K concentration of 109.0 +/- 9.1 mM, and an intracellular Na concentration of 47.0 +/- 13.4mM.) The incorporation of [14C]uridine also sensitively reflects cellular damage.
36.4 +/测量了各种细胞参数,以评估它们作为分离细胞活力标准的有用性。台盼蓝染色性和乳酸脱氢酶释放仅表明细胞受到严重的不可逆损伤。内源性呼吸甚至ATP/ADP比值都不是细胞活力的敏感标准。在细胞衰老过程中,即使膜电位、细胞内钾浓度和腺嘌呤核苷酸含量大幅下降,ATP/ADP比值仍保持较高水平。一个敏感且易于进行的测试是用1mM琥珀酸刺激细胞呼吸。只有受损的质膜才会使琥珀酸以足以刺激呼吸的速率渗透。膜电位以及细胞内钠和钾浓度是细胞活力最敏感的标准,因为它们表明衰老过程中最早出现的变化。(对于新鲜分离的细胞,我们发现膜电位为36.4 ± 3.4 mV [n = 5],细胞内钾浓度为109.0 ± 9.1 mM,细胞内钠浓度为47.0 ± 13.4 mM。)[14C]尿苷的掺入也能敏感地反映细胞损伤。