ISSN:
0006-3592
Keywords:
hubridoma
;
antibody
;
osmolarty
;
aminoacide uptake
;
Na+-dependent transport system
;
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
It has previously been found tht hybridoma cells undr hyuerosmotic stress produce higher amounts of antibody. This study indentified the cellular processes and mechanisms that occur during this event. In studies fo hybridomas adpated toosmolarities ranging between 300 and 450 mOsm (uusing NaCl), antibody production increased to a saturation level while cell growth decreased progressively. At 500 mOsm, lower, cell numbers and markedly decreaased productivity resulted. Sucrose and KCl were found to induce similar trends, except to different extents.Several important change in cellulaes in cellular responses were onsserved. Elevation of osmnolarity with NaCl from 300 to 350 mOsm causes an increase of zwiterionic amino acid upatake, which, occurredvia Na+-dependent transport systems. In particuar, systedm A was enhanced by 1.86-fold, but noenhancement was observed for Na+-independent transport systems, In addition, amino acids reactive with Na+-dependent transport systems were onserved to be abundant within osmotically stressed hybridomas in the middle and dlate exponentoial statges. Sucroses ans Kcl caused similar uptake effects, but to a laeeser degree, as long as sodium ions were present in solution.Specific consumption rates fo glucose and glutamine incresase by 19% and 20%, respectively, under high osmolarity treatment. Thewse increases were confirmed by the 5% to 10% increase in cellular metabolic acitivity. At 350 mOsm, growth rate was slower, compared with the 300-mOsm culture, which was reflected by thelower DNA conetr4ation. Stressed cultures contained enhanced leyls of tatal RNA content could in turn increase the translation rates of proteins. This was reflected in the accumulation of both dry cell weight and total cellular protein at linear rates of 0.42 μG/106 cells/mOsm and 0.21 μg/106 cells/mOmsm, respectively, with increasing osmolarty between 300 and 450 mOsm.Overall, hybridoms increased their metabolic activities and amino acids uptake via the Na+-dependent symports to compensate for teh osmotically elevated external environment. These effects contribute directly and indirectly tothe increased cell mass consisting of a larger pool of amono acids, RNA, cellular proteins, and seecreted antibody produt. © 1995 John Wiley & Sons, Inc.
Additional Material:
6 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/bit.260460605
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