Yeast hydrolysate as a low-cost additive to serum-free medium for the production of human thrombopoietin in suspension cultures of Chinese hamster ovary cells.
To enhance the performance of a serum-free medium (SFM) for human thrombopoietin (hTPO) production in suspension cultures of recombinant Chinese hamster ovary (rCHO) cells, several low-cost hydrolysates such as yeast hydrolysate (YH), soy hydrolysate, wheat gluten hydrolysate and rice hydrolysate were tested as medium additives. Among various hydrolysates tested, the positive effect of YH on hTPO production was most significant. When 5 g l(-1) YH was added to SFM, the maximum hTPO concentration in batch culture was 40.41 microg ml(-1), which is 11.5 times higher than that in SFM without YH supplementation. This enhanced hTPO production in YH-supplemented SFM was obtained by the combined effect of enhanced q(hTPO) (the specific rate of hTPO production). The supplementation of YH in SFM increased q(hTPO) by 294% and extended culture longevity by >2 days if the culture was terminated at a cell viability of 50%. Furthermore, cell viability throughout the culture using YH-supplemented SFM was higher than that using any other hydrolysate-supplemented SFM tested, thereby minimizing degradation of hTPO susceptible to proteolytic degradation. In addition, YH supplementation did not affect in vivo biological activity of hTPO. Taken together, the results obtained demonstrate the potential of YH as a medium additive for hTPO production in serum-free suspension cultures of rCHO cells.
Sung
Y H
YH
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Kusong-Dong, Yusong-Gu, 305-701 Daejon, Korea..
Lim
S W
SW
Chung
J Y
JY
Lee
G M
GM
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free / Q000737:chemistry*; D005656:Fungal Proteins; D005983:Glutens; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D012275:Oryza; D010940:Plant Proteins; D010976:Platelet Count; D011492:Protein Hydrolysates; D011994:Recombinant Proteins / Q000096:biosynthesis; D030262:Soybean Proteins; D013926:Thrombopoietin / Q000096:biosynthesis*; D013997:Time Factors
D000596:Amino Acids; D016895:Culture Media, Serum-Free; D005656:Fungal Proteins; D010940:Plant Proteins; D011492:Protein Hydrolysates; D011994:Recombinant Proteins; D030262:Soybean Proteins; D005983:Glutens; D013926:Thrombopoietin; D007770:L-Lactate Dehydrogenase