Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids

Bradley P. Weegman, Ahmad Essawy, Peter Nash, Alexandra L. Carlson, Kristin J. Voltzke, Zhaohui Geng, Marjan Jahani, Benjamin B. Becker, Klearchos K. Papas, Meri T. Firpo

Research output: Contribution to journalArticle

  • 1 Citations

Abstract

In this demonstration, spheroids formed from the β-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.

Original languageEnglish (US)
Article numbere52224
JournalJournal of Visualized Experiments
Volume2016
Issue number115
DOIs
StatePublished - Sep 25 2016

Fingerprint

Glucose
Nutrients
Cells
Cell growth
Bioreactors
Demonstrations

Keywords

  • Bioengineering
  • Bioreactor
  • Continuous feeding
  • Culture
  • Expansion
  • Glucose
  • Issue 115
  • Nutrient regulation
  • Spheroid
  • Stirred suspension bioreactor

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

MeSH PubMed subject areas

  • Journal Article
  • Video-Audio Media

Cite this

Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids. / Weegman, Bradley P.; Essawy, Ahmad; Nash, Peter; Carlson, Alexandra L.; Voltzke, Kristin J.; Geng, Zhaohui; Jahani, Marjan; Becker, Benjamin B.; Papas, Klearchos K.; Firpo, Meri T.

In: Journal of Visualized Experiments, Vol. 2016, No. 115, e52224, 25.09.2016.

Research output: Contribution to journalArticle

Weegman, BP, Essawy, A, Nash, P, Carlson, AL, Voltzke, KJ, Geng, Z, Jahani, M, Becker, BB, Papas, KK & Firpo, MT 2016, 'Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids' Journal of Visualized Experiments, vol 2016, no. 115, e52224. DOI: 10.3791/52224
Weegman BP, Essawy A, Nash P, Carlson AL, Voltzke KJ, Geng Z et al. Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids. Journal of Visualized Experiments. 2016 Sep 25;2016(115). e52224. Available from, DOI: 10.3791/52224

Weegman, Bradley P.; Essawy, Ahmad; Nash, Peter; Carlson, Alexandra L.; Voltzke, Kristin J.; Geng, Zhaohui; Jahani, Marjan; Becker, Benjamin B.; Papas, Klearchos K.; Firpo, Meri T. / Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids.

In: Journal of Visualized Experiments, Vol. 2016, No. 115, e52224, 25.09.2016.

Research output: Contribution to journalArticle

@article{9273d27647a54756b61e2bd9638978eb,
title = "Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids",
abstract = "In this demonstration, spheroids formed from the β-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.",
keywords = "Bioengineering, Bioreactor, Continuous feeding, Culture, Expansion, Glucose, Issue 115, Nutrient regulation, Spheroid, Stirred suspension bioreactor",
author = "Weegman, {Bradley P.} and Ahmad Essawy and Peter Nash and Carlson, {Alexandra L.} and Voltzke, {Kristin J.} and Zhaohui Geng and Marjan Jahani and Becker, {Benjamin B.} and Papas, {Klearchos K.} and Firpo, {Meri T.}",
year = "2016",
month = "9",
doi = "10.3791/52224",
volume = "2016",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "115",

}

TY - JOUR

T1 - Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids

AU - Weegman,Bradley P.

AU - Essawy,Ahmad

AU - Nash,Peter

AU - Carlson,Alexandra L.

AU - Voltzke,Kristin J.

AU - Geng,Zhaohui

AU - Jahani,Marjan

AU - Becker,Benjamin B.

AU - Papas,Klearchos K.

AU - Firpo,Meri T.

PY - 2016/9/25

Y1 - 2016/9/25

N2 - In this demonstration, spheroids formed from the β-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.

AB - In this demonstration, spheroids formed from the β-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.

KW - Bioengineering

KW - Bioreactor

KW - Continuous feeding

KW - Culture

KW - Expansion

KW - Glucose

KW - Issue 115

KW - Nutrient regulation

KW - Spheroid

KW - Stirred suspension bioreactor

UR - http://www.scopus.com/inward/record.url?scp=84989318323&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84989318323&partnerID=8YFLogxK

U2 - 10.3791/52224

DO - 10.3791/52224

M3 - Article

VL - 2016

JO - Journal of Visualized Experiments

T2 - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 115

M1 - e52224

ER -