Barker G, Simmons N L
Q J Exp Physiol. 1981 Jan;66(1):61-72. doi: 10.1113/expphysiol.1981.sp002529.
Cultured monolayers of Madin-Derby canine kidney (MDCK) cells grown upon permeable filter supports display many features of in vivo epithelia. Measurements of transmonolayer resistance, open-circuit p.d., dilution and bi-ionic p.d.s have been made of MDCK monolayers mounted in Ussing-type chamber (0.75 cm window radius). Previously reported values of transmonolayer resistance of 100 omega cm2 (Cereijido, Robbins, Dolan, Rotunno & Sabatini, 1978; Misfeldt, Hamamoto & Pitelka, 1976) indicate a 'leaky' epithelium. The major finding of this work is that MDCK monolayers can also display values of transmonolayer resistance similar to 'tight' epithelia (mean value = 4116 omega cm2). The reason for such differences is the existence of separate strains of MDCK cells. High-resistance monolayers originate from cell stocks of sixty serial passages whilst low-resistance monolayers originate from cell stocks of 109 serial passages. Measurements of transmonolayer dilution, and bi-ionic p.d.s together with Na ion tracer fluxes and electron microscope observations of La3+ penetration through the apical tight junctions, substantiate the finding of high-resistance properties in MDCK monolayers and confirm the existence of two separate MDCK cell strains displaying entirely different physiological properties. Differences extend to cellular morphology and cellular volume in the two strains of MDCK cells.
在可渗透滤膜支持物上生长的Madin-Darby犬肾(MDCK)细胞培养单层显示出许多体内上皮细胞的特征。对安装在Ussing型小室(窗口半径0.75厘米)中的MDCK单层进行了跨单层电阻、开路电位差、稀释和双离子电位差的测量。先前报道的跨单层电阻值为100Ω·cm²(Cereijido、Robbins、Dolan、Rotunno和Sabatini,1978年;Misfeldt、Hamamoto和Pitelka,1976年),表明是一种“渗漏性”上皮。这项工作的主要发现是,MDCK单层也可以显示出与“紧密性”上皮相似的跨单层电阻值(平均值 = 4116Ω·cm²)。这种差异的原因是存在不同的MDCK细胞株。高电阻单层源自60代连续传代的细胞株,而低电阻单层源自109代连续传代的细胞株。跨单层稀释和双离子电位差的测量,以及Na离子示踪剂通量的测量和La³⁺通过顶端紧密连接渗透的电子显微镜观察,证实了MDCK单层具有高电阻特性的发现,并确认存在两种显示出完全不同生理特性的独立MDCK细胞株。差异还延伸到两种MDCK细胞株的细胞形态和细胞体积。