EurekaMag.com logo
+ Translate

Preparation of individual human diploid fibroblasts and study of ion transport


, : Preparation of individual human diploid fibroblasts and study of ion transport. American Journal of Physiology 248(1 Pt 1): C154-C164

A method for analyzing individual mammalian cells with electron probe microanalysis has been developed using human diploid fibroblasts. Cells were grown on the same support that is used for experimental manipulations and analysis. Steady-state cation and anion concentrations and kinetic processes during experimental perturbations could be measured on populations of less than 1,000 cells. Human diploid fibroblasts in normal tissue culture medium had the following intracellular concentrations (in mM): K, 168; Na, 25.0; Cl, 51.2; P, 84.1; S, 16.5; Ca, 6.04; and Mg, 10.0. The ratios of K to Na were equivalent when measured in the nuclear or cytoplasmic area of the cells. Serum in the incubation medium was found to increase the cellular effective permeability to Na by a factor of 2.5, while leaving the effective permeability to K unchanged. When returned to control medium after 7 h of incubation in K-free medium, the cells recovered normal K/Na in less than 1 h. In some experiments the coupling ratio of the ouabain-inhibitable cellular transport of Na to K was 3:2 and the ratio of Cl to K was 1:2. The sum of intracellular content (Na + K) (an estimate of cellular volume) did not change when the cells were placed in K-free medium and increased by less than 30% after ouabain treatment. After 5-7 h of ouabain treatment or of incubation in K-free medium, long after the intracellular K had been replaced by Na, the cellular chloride content had not changed significantly.

(PDF 0-2 workdays service)

Accession: 006173314

PMID: 3966540

Submit PDF Full Text: Here


Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:
Close
Close

Related references

Rao M.V.N., 1985: Differences in the response of non dividing human diploid fibroblasts to inducers of dna synthesis generated by cycling human diploid fibroblasts and hela cells. The ability of non-dividing WI-38 cells from different passages to respond to inducers of DNA synthesis generated by other WI-38 cells in G1 or S and HeLa cells in S was studied. The non-dividing cells are refractory to inducers of DNA synthesis g...

Terekhov S.M.; Getsadze K.A.; Grinberg K.N., 1984: Individual variability of proliferative potential of human diploid fibroblasts in vitro. The rate of individual variability of proliferative potential determined by the percentage of colonies containing 16 and more cells (the cloning efficiency) was studied in 16 embyronic and 15 postnatal strains of human diploid fibroblasts. These d...

Cook K.R.; Pool T.B., 1984: Detection of collagen messenger rna production by individual human diploid fibroblasts. Journal of Cell Biology 99(4 PART 2): 406A

Terekhov S.M.; Getsadze K.A.; Grinberg K.N., 1984: Individual variation in proliferative potential of human diploid fibroblasts in vitro. Bulletin of Experimental Biology & Medicine 97(4): 479-480

Terekhov, S.M.; Getsadze, K.A.; Grinberg, K.N., 1984: Individual variability in the proliferative potential of human diploid fibroblasts in vitro. The rate of individual variability of proliferative potential determined by the percentage of colonies containing 16 and more cells (the cloning efficiency) was studied 16 embryonic and 15 postnatal strains of human diploid fibroblasts. It is high...

Dikomey, E.; Borgmann, K.; Brammer, I.; Kasten-Pisula, U., 2003: Molecular mechanisms of individual radiosensitivity studied in normal diploid human fibroblasts. The molecular mechanisms of individual radiosensitivity were studied in normal diploid human fibroblasts. For fibroblasts irradiated with X-rays in G1-phase the individual radiosensitivity was shown to be correlated with the extent of double-stran...

Corriat, A.; Moatti, N.; Lemonnier, A., 1983: Transport of L-histidine by human diploid fibroblasts in culture. The transport of L-histidine has been characterized in skin derived diploid human fibroblasts, cultured under strictly controlled conditions. The transport measurements were made on cells grown to subconfluency after 60 to 90 min timed preincubati...

Augenlicht, L.H.; Baserga, R., 1973: Preparation and partial fractionation of nonhistone chromosomal proteins from human diploid fibroblasts. Archives of Biochemistry and Biophysics 158(1): 89-96

Germinario, R.J.; Rockman, H.; Oliveira, M.; Manuel, S.; Taylor, M., 1982: Regulation of sugar transport in cultured diploid human skin fibroblasts. The regulation of hexose transport under glucose-starvation conditions was studied in cultured human skin fibroblasts. Glucose starvation enhanced the transport of 2-DG and 3-0-methyl-D-glucose (3-OMG) but not of L-glucose. Glucose-starvation enha...

Bannai, S.; Kitamura, E., 1980: Transport interaction of L-cystine and L-glutamate in human diploid fibroblasts in culture. Journal of Biological Chemistry 255(6): 2372-2376