An engineered approach to stem cell culture: Automating the decision process for real-time adaptive subculture of stem cells

Dai Fei Elmer Ker, Lee E. Weiss, Silvina N. Junkers, Mei Chen, Zhaozheng Yin, Michael F. Sandbothe, Seung Il Huh, Sungeun Eom, Ryoma Bise, Elvira Osuna-Highley, Takeo Kanade, Phil G. Campbell

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Current cell culture practices are dependent upon human operators and remain laborious and highly subjective, resulting in large variations and inconsistent outcomes, especially when using visual assessments of cell confluency to determine the appropriate time to subculture cells. Although efforts to automate cell culture with robotic systems are underway, the majority of such systems still require human intervention to determine when to subculture. Thus, it is necessary to accurately and objectively determine the appropriate time for cell passaging. Optimal stem cell culturing that maintains cell pluripotency while maximizing cell yields will be especially important for efficient, cost-effective stem cell-based therapies. Toward this goal we developed a real-time computer vision-based system that monitors the degree of cell confluency with a precision of 0.791±0.031 and recall of 0.559±0.043. The system consists of an automated phase-contrast time-lapse microscope and a server. Multiple dishes are sequentially imaged and the data is uploaded to the server that performs computer vision processing, predicts when cells will exceed a pre-defined threshold for optimal cell confluency, and provides a Web-based interface for remote cell culture monitoring. Human operators are also notified via text messaging and e-mail 4 hours prior to reaching this threshold and immediately upon reaching this threshold. This system was successfully used to direct the expansion of a paradigm stem cell population, C2C12 cells. Computer-directed and human-directed control subcultures required 3 serial cultures to achieve the theoretical target cell yield of 50 million C2C12 cells and showed no difference for myogenic and osteogenic differentiation. This automated vision-based system has potential as a tool toward adaptive real-time control of subculturing, cell culture optimization and quality assurance/quality control, and it could be integrated with current and developing robotic cell cultures systems to achieve complete automation.

Original languageEnglish
Article numbere27672
JournalPloS one
Volume6
Issue number11
DOIs
Publication statusPublished - Nov 16 2011

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Stem cells
Cell culture
stem cells
cell culture
Stem Cells
Cell Culture Techniques
Computer vision
cells
Robotics
Servers
Text messaging
Real time control
computer vision
Quality assurance
Quality control
quality control
Microscopes
Automation
Text Messaging
Monitoring

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Ker, D. F. E., Weiss, L. E., Junkers, S. N., Chen, M., Yin, Z., Sandbothe, M. F., ... Campbell, P. G. (2011). An engineered approach to stem cell culture: Automating the decision process for real-time adaptive subculture of stem cells. PloS one, 6(11), [e27672]. https://doi.org/10.1371/journal.pone.0027672

An engineered approach to stem cell culture : Automating the decision process for real-time adaptive subculture of stem cells. / Ker, Dai Fei Elmer; Weiss, Lee E.; Junkers, Silvina N.; Chen, Mei; Yin, Zhaozheng; Sandbothe, Michael F.; Huh, Seung Il; Eom, Sungeun; Bise, Ryoma; Osuna-Highley, Elvira; Kanade, Takeo; Campbell, Phil G.

In: PloS one, Vol. 6, No. 11, e27672, 16.11.2011.

Research output: Contribution to journalArticle

Ker, DFE, Weiss, LE, Junkers, SN, Chen, M, Yin, Z, Sandbothe, MF, Huh, SI, Eom, S, Bise, R, Osuna-Highley, E, Kanade, T & Campbell, PG 2011, 'An engineered approach to stem cell culture: Automating the decision process for real-time adaptive subculture of stem cells', PloS one, vol. 6, no. 11, e27672. https://doi.org/10.1371/journal.pone.0027672
Ker, Dai Fei Elmer ; Weiss, Lee E. ; Junkers, Silvina N. ; Chen, Mei ; Yin, Zhaozheng ; Sandbothe, Michael F. ; Huh, Seung Il ; Eom, Sungeun ; Bise, Ryoma ; Osuna-Highley, Elvira ; Kanade, Takeo ; Campbell, Phil G. / An engineered approach to stem cell culture : Automating the decision process for real-time adaptive subculture of stem cells. In: PloS one. 2011 ; Vol. 6, No. 11.
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