Determination of Minimum Inhibitory Concentration (MIC) of Hygromycin B in CHO cells The minimum inhibitory concentration determination
Iranian Journal of Pharmaceutical Sciences,
Vol. 15 Núm. 3 (2019),
1 July 2019
,
Página 1-6
https://doi.org/10.22037/ijps.v15.40472
Resumen
Chinese hamster ovary (CHO) cells are considered as the most commonly used host for industrial manufacturing of therapeutic proteins. The aim of this study was to evaluate the minimum inhibitory concentration (MIC) of hygromycin B in both CHO-DG44 and CHO-S cells since hygromycin B resistance cassette can be used for future selection of gene expression in CHO cells. The minimum inhibitory concentration was determined by the microdilution method. In this test, CHO cells were subjected to a range of hygromycin B concentrations for a defined period of time. Moreover, CHO cells were scaled up with no addition of fresh culture medium during the growth–culture cycle to investigate their growth. The MIC for hygromycin B in CHO-DG44 cells was 75 μg/ml and for CHO-S cells was 50 μg/ml. Thus, it has been indicated that CHO DG44 cell line could be more effective for stable cell line development than CHO-S cell line.
- Cell culture
- Chinese hamster ovary cells
- CHO-DG44
- CHO-S
- MIC
- hygromycin B
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[2] Kim BJ, Zhao T, Young L, Zhou P, Shuler ML. Batch, fed‐batch, and microcarrier cultures with CHO cell lines in a pressure‐cycle driven miniaturized bioreactor. Biotechnol. Bioeng. (2012) 109 (1): 137-45.
[3] Delrue I, Pan Q, Baczmanska AK, Callens BW, Verdoodt LL. Determination of the Selection Capacity of Antibiotics for Gene Selection. Biotechnol. J. (2018): 1700747.
[4] McGaha SM, Champney WS. Hygromycin B inhibition of protein synthesis and ribosome biogenesis in Escherichia coli. Antimicrob. Agents Ch. (2007) 51 (2): 591-6.
[5] Ganoza MC, Kiel MC. A ribosomal ATPase is a target for hygromycin B inhibition on Escherichia coli ribosomes. Antimicrob. Agents Ch. (2001) 45 (10): 2813-9.
[6] Brodersen DE, Clemons Jr WM, Carter AP, Morgan-Warren RJ, Wimberly BT, Ramakrishnan V. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell J. (2000) 103 (7): 1143-54.
[7] Moazed D, Noller HF. Interaction of antibiotics with functional sites in 16S ribosomal RNA. Nature. 1987;327(6121):389.
[8] Goswami J, Sinskey A, Steller H, Stephanopoulos G, Wang D. Apoptosis in batch cultures of Chinese hamster ovary cells. Biotechnol. Bioeng. (1999) 62 (6): 632-40.
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