Cargando…
An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells
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 t...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218005/ https://www.ncbi.nlm.nih.gov/pubmed/22110715 http://dx.doi.org/10.1371/journal.pone.0027672 |
_version_ | 1782216651537645568 |
---|---|
author | 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. |
author_facet | 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. |
author_sort | Ker, Dai Fei Elmer |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3218005 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-32180052011-11-21 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. PLoS One Research Article 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. Public Library of Science 2011-11-16 /pmc/articles/PMC3218005/ /pubmed/22110715 http://dx.doi.org/10.1371/journal.pone.0027672 Text en This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication. https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. |
spellingShingle | Research Article 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 |
title | An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells |
title_full | An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells |
title_fullStr | An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells |
title_full_unstemmed | An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells |
title_short | An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells |
title_sort | engineered approach to stem cell culture: automating the decision process for real-time adaptive subculture of stem cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218005/ https://www.ncbi.nlm.nih.gov/pubmed/22110715 http://dx.doi.org/10.1371/journal.pone.0027672 |
work_keys_str_mv | AT kerdaifeielmer anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT weissleee anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT junkerssilvinan anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT chenmei anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT yinzhaozheng anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT sandbothemichaelf anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT huhseungil anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT eomsungeun anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT biseryoma anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT osunahighleyelvira anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT kanadetakeo anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT campbellphilg anengineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT kerdaifeielmer engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT weissleee engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT junkerssilvinan engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT chenmei engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT yinzhaozheng engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT sandbothemichaelf engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT huhseungil engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT eomsungeun engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT biseryoma engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT osunahighleyelvira engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT kanadetakeo engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells AT campbellphilg engineeredapproachtostemcellcultureautomatingthedecisionprocessforrealtimeadaptivesubcultureofstemcells |