An improved approach to freeze-fracture morphology of monolayer cell cultures

1998 ◽  
Vol 82 (1) ◽  
pp. 97-103 ◽  
Author(s):  
Elisabeth Bugnard ◽  
Patrick Sors ◽  
Alain Bloc ◽  
Françoise Loctin ◽  
Yves Dunant
1992 ◽  
Vol 22 (3) ◽  
pp. 301-305 ◽  
Author(s):  
John H. Todd ◽  
Donald A. Sens ◽  
Mary Ann Sens ◽  
Debra Hazen-Martin

1988 ◽  
Vol 90 (1) ◽  
pp. 99-104
Author(s):  
Z. Szallasi ◽  
A. Szallasi ◽  
F. Bojan ◽  
I. Zs-Nagy

Swiss/3T3 cell cultures were harvested with 0.05% collagenase and after centrifugation the pellet was prepared by the freeze-fracture/freeze-drying (FFFD) method for bulk-specimen X-ray microanalysis. Time-dependent variations in the intracellular monovalent elemental concentrations (Na+, K+ and Cl-) as well as of the Na+/K+ ratio were followed for 120 min subsequent to harvesting. The quantitative measurements revealed a very considerable increase in the intracellular Na+ and Cl- accompanied by a decrease in the K+ concentration as soon as 5 min after harvesting. The Na+/K+ ratio had increased by this time to about 1.5 on average. These changes indicate a sustained depolarization of the cell membrane. During the first 60 min this depolarization tended to normalize as demonstrated by an exponential decrease in the intracellular Na+ and Cl- and an increase in the K+ content involving a decrease in the Na+/K+ ratio. The total intracellular monovalent ion concentration remained almost constant during this post-harvesting period. These results suggest that harvesting represents a serious depolarizing stimulus to the cells, the consequences of which are restored only after 1–2h. These alterations should be taken into consideration during various experimental designs when using anchorage-dependent cell cultures.


2010 ◽  
Vol 152-153 ◽  
pp. 924-930
Author(s):  
Mei Li ◽  
Zhi Qiang Li ◽  
Jun Xu ◽  
Dun Sheng Wei ◽  
Hong Wei Zhu ◽  
...  

Modified Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] was prepared by melt reactive blending P(3HB-co-4HB) with chain extenders (ADR-4367). Thermal transitions, spherulitic and freeze-fracture morphology, mechanical and rheological properties of the chain extended bio-materials were investigated. The results show that glass transition temperatures and crystallization temperatures increase, crystallinity and diameters of spherulites decrease, there are amorphous parts dispersed among the crystalline phase, and the bio-materials transfer from brittleness to toughness and ductility. Steady shear viscosity of the modified P(3HB-co-4HB) increases by about one order of magnitude, melts of the modified P(3HB-co-4HB) behave more viscoelasticity by storage modulus and loss modulus correlated with oscillatory shear frequency. Addition of ADR-4367 with contents of 4~6 wt% in the blends is enough to branching and coupling the co-polymer chains and brings remarkable effect on improving mechanical properties, steady shear viscosity and viscoelasticity.


1991 ◽  
Vol 131 (2) ◽  
pp. 287-293 ◽  
Author(s):  
P. Y. D. Wong ◽  
C. N. Uchendu

ABSTRACT Monolayer cell cultures formed from the rat cauda epididymidis exhibited renin-like and angiotensin-converting enzyme (ACE) activities and contained immunoreactive angiotensin I (AI) and angiotensin II (AII). Renin-like activity, determined indirectly by radioimmunoassay of generated AI at a near-neutral pH of 6·0, was demonstrated in the cell lysate but was almost undetectable in the serum-free cell culture medium, suggesting that renin expression in epididymal cells is an intracellular phenomenon. In contrast, both AI and AII were detected in the cell lysate and cell culture medium. The level of AI was enhanced by pretreating the cells with the ACE inhibitor captopril (100 nmol/l). Incubating the cell monolayers with thoroughly washed sperm cells obtained from the intact cauda epididymides of rats increase (P < 0·01) the AII content of the cell culture medium, with a parallel decline (P < 0·01) in the AI concentration. However, adrenaline (0·23 μmol/l), which was found to stimulate electrogenic anion secretion by cell monolayers grown on pervious supports, was without effect on the renin-like activity or concentration of angiotensins. The ACE activity in cells was confirmed by its strong dependence on chloride ion and its susceptibility to inhibition by captopril (100 nmol/l). Enzyme activity was significantly (P < 0·005) higher in the culture medium than in the cell lysate and cell membrane fragments. Angiotensinogen, which is obligatory for an intrinsic renin-angiotensin system, is present in epididymal cells. Presumably, it is synthesized and processed in the cell cytosol by intracellular renin. These findings in cultured cells provide further evidence for a local renin–angiotensin system in epididymal tissue and suggest a possible role of endogenous AII in the regulation of epididymal function. Journal of Endocrinology (1991) 131, 287–293


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