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,Seungil Huh,Sungeun Eom,Ryoma Bise,Elvira Osuna-Highley,Takeo Kanade,Phil G. Campbell +11 more
TLDR
An automated vision-based system that monitors the degree of cell confluency and could be integrated with current and developing robotic cell cultures systems to achieve complete automation has potential as a tool toward adaptive real-time control of subculturing, cell culture optimization and quality assurance/quality control.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.read more
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Journal ArticleDOI
Automated method for the rapid and precise estimation of adherent cell culture characteristics from phase contrast microscopy images
Nicolas Jaccard,Lewis D. Griffin,Ana Keser,Rhys J. Macown,Alexandre Super,Farlan S. Veraitch,Nicolas Szita +6 more
TL;DR: This work presents an image‐processing approach that accurately detects cellular objects in PCM images through a combination of local contrast thresholding and post hoc correction of halo artifacts, and created a software toolbox (PHANTAST) that bundles all the algorithms and provides an easy to use graphical user interface.
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Cell motion predicts human epidermal stemness
Daisuke Nanba,Fujio Toki,Sota Tate,Matome Imai,Natsuki Matsushita,Ken Shiraishi,Koji Sayama,Hiroshi Toki,Shigeki Higashiyama,Yann Barrandon,Yann Barrandon +10 more
TL;DR: Keratinocyte stem cell colonies can be identified by analyzing cell motion, an emergent stem cell property, which indicates whether the colony is occupied by a single cell or several cells.
Journal ArticleDOI
Automated and Online Characterization of Adherent Cell Culture Growth in a Microfabricated Bioreactor
Nicolas Jaccard,Rhys J. Macown,Alexandre Super,Lewis D. Griffin,Farlan S. Veraitch,Nicolas Szita +5 more
TL;DR: A microfabricated bioreactor that enables a high degree of control on the microenvironment of the cells while remaining compatible with standard cell culture protocols is described and its integration with automated image-processing capabilities is described, allowing the continuous monitoring of key cell culture characteristics.
Journal ArticleDOI
Selection and tuning of a fast and simple phase-contrast microscopy image segmentation algorithm for measuring myoblast growth kinetics in an automated manner.
TL;DR: A simple and fast algorithm to compute the cell-covered surface (degree of confluence) in phase-contrast microscopy images is developed based on a range filter of a specified size, a minimum range threshold, and a minimum object size threshold.
Journal ArticleDOI
Predicting in vitro human mesenchymal stromal cell expansion based on individual donor characteristics using machine learning.
Mohammad Mehrian,Mohammad Mehrian,Toon Lambrechts,Marina Marechal,Frank P. Luyten,Ioannis Papantoniou,Liesbet Geris,Liesbet Geris +7 more
TL;DR: A predictive computational model describing in vitro cell expansion dynamics based on individual donor characteristics is introduced, an approach that could greatly assist toward automation of a cell expansion culture process.
References
More filters
Journal ArticleDOI
Rapid Hematopoietic Recovery After Coinfusion of Autologous-Blood Stem Cells and Culture-Expanded Marrow Mesenchymal Stem Cells in Advanced Breast Cancer Patients Receiving High-Dose Chemotherapy
Omer N. Koc,Stanton L. Gerson,Brenda W. Cooper,Stephanie M. Dyhouse,Stephen E. Haynesworth,Arnold I. Caplan,Hillard M. Lazarus +6 more
TL;DR: It is found that autologous MSC infusion at the time of PBPC transplantation is feasible and safe, and the observed rapid hematopoietic recovery suggests that M SC infusion after myeloablative therapy may have a positive impact on hematoiesis and should be tested in randomized trials.
Journal ArticleDOI
Plasticity of the differentiated state.
Helen M. Blau,Grace K. Pavlath,Edna C. Hardeman,Choy-Pik Chiu,Laura Silberstein,Steven G. Webster,S C Miller,Cecelia Webster +7 more
TL;DR: Results show that the expression of genes in the nuclei of differentiated cells is remarkably plastic and susceptible to modulation by the cytoplasm, and the isolation of the genes encoding the tissue-specific trans-acting regulators responsible for muscle gene activation should now be possible.
Book
Fundamentals of Light Microscopy and Electronic Imaging
TL;DR: In this article, the authors discuss the fundamental principles of light microscopy, including light and color illuminators, filters, and isolation of specific wavelengths of light and reflectors.
Journal ArticleDOI
BMP2 Regulates Osterix through Msx2 and Runx2 during Osteoblast Differentiation
Takuma Matsubara,Kumiko Kida,Akira Yamaguchi,Kenji Hata,Fumitaka Ichida,Hiroko Meguro,Hiroyuki Aburatani,Riko Nishimura,Toshiyuki Yoneda +8 more
TL;DR: Osterix is regulated via both Runx2-dependent and -independent mechanisms, and that Osterix controls osteoblast differentiation, at least in part, by regulating the expression of genes not controlled by Runx 2.
DissertationDOI
Efficient Algorithms for Sorting and Synchronization
TL;DR: This thesis presents efficient algorithms for internal and external parallel sorting and remote data update and examines a number of related algorithms for text compression, differencing and incremental backup.