Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells. (23rd April 2021)

• Record Type: Journal Article
• Authors: Wu, Suyang; Rish, Adam J.; Skomo, Alec; Zhao, Yuxiang; Drennen, James K.; Anderson, Carl A.
• Is Part Of: Biotechnology progress. Volume 37:Number 4(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-04-23
• Subjects: CHO cells -- definitive screening design -- integrin -- serum‐free/suspension adaptation; Biotechnology -- Periodicals; Food industry and trade -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/btpr.3154
• Languages: English
• ISSNs: 8756-7938

Abstract

Abstract: The biopharmaceutical industry prefers to culture the mammalian cells in suspension with a serum‐free media (SFM) due to improved productivity and process consistency. However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require adaptation from adherence in CM to suspension culture in SFM. This work proposes an adaptation method that includes media supplementation during the adaption of Chinese hamster ovary cells. As a result, the adaptation was accelerated compared to the traditional repetitive subculturing. Ca 2+ /Mg 2+ supplementation significantly reduced the doubling time compared to the adaptation without supplementation during the adaptation of adherent cells from 100% CM to 75% CM ( p < 0.05). Furthermore, a definitive screening design (DSD) was applied to select essential nutrients during the adaptation from 10% CM to 0% CM. The main effects of Ca 2+ and Dulbecco's modified essential medium (DMEM) were found significant to both viable cell density and viability at harvest. Additionally, the interaction term between Ca 2+ and DMEM was found significant, which highlights the ability of DSD to capture interaction terms. Eventually, the media supplementation method resulted in adaptation SFM in 27 days, compared to the previously reported 66 days. Additionally, the membrane surface integrin expression was found significantly decreased when adherent cells were adapted to suspension. Moreover, the Ca 2+ /Mg 2+ supplementation correlated with faster integrin recovery after trypsinization. However, faster integrin recovery did not contribute to the accelerated cell growth when subculturing from 100% CM to 75% CM.