Automated image analysis detects aging in clinical-grade mesenchymal stromal cell cultures

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http://hdl.handle.net/10138/231830

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Oja , S , Komulainen , P , Penttilä , A , Nystedt , J & Korhonen , M 2018 , ' Automated image analysis detects aging in clinical-grade mesenchymal stromal cell cultures ' , Stem Cell Research & Therapy , vol. 9 , 6 . https://doi.org/10.1186/s13287-017-0740-x

Title: Automated image analysis detects aging in clinical-grade mesenchymal stromal cell cultures
Author: Oja, S.; Komulainen, P.; Penttilä, A.; Nystedt, J.; Korhonen, M.
Other contributor: University of Helsinki, Department of Physics
University of Helsinki, Children's Hospital


Date: 2018-01-10
Language: eng
Number of pages: 13
Belongs to series: Stem Cell Research & Therapy
ISSN: 1757-6512
DOI: https://doi.org/10.1186/s13287-017-0740-x
URI: http://hdl.handle.net/10138/231830
Abstract: Background: Senescent cells are undesirable in cell therapy products due to reduced therapeutic activity and risk of aberrant cellular effects, and methods for assessing senescence are needed. Early-passage mesenchymal stromal cells (MSCs) are known to be small and spindle-shaped but become enlarged upon cell aging. Indeed, cell morphology is routinely evaluated during MSC production using subjective methods. We have therefore explored the possibility of utilizing automated imaging-based analysis of cell morphology in clinical cell manufacturing. Methods: An imaging system was adopted for analyzing changes in cell morphology of bone marrow-derived MSCs during long-term culture. Cells taken from the cultures at the desired passages were plated at low density for imaging, representing morphological changes observed in the clinical-grade cultures. The manifestations of aging and onset of senescence were monitored by population doubling numbers, expression of p16(INK4)a and p21(Cip1/Waf1), beta-galactosidase activity, and telomeric terminal restriction fragment analysis. Results: Cell area was the most statistically significant and practical parameter for describing morphological changes, correlating with biochemical senescence markers. MSCs from passages 1 (p1) and 3 (p3) were remarkably uniform in size, with cell areas between 1800 and 2500 mu m(2). At p5 the cells began to enlarge resulting in a 4.8-fold increase at p6-9 as compared to p1. The expression of p16(INK4a) and activity of beta-galactosidase had a strong correlation with the increase in cell area, whereas the expression of p21(Cip1/Waf1) reached its maximum at the onset of growth arrest and subsequently decreased. Mean telomere length shortened at an apparently constant rate during culture, from 8.2 +/- 0.3 kbp at p1, reaching 6.08 +/- 0.6 kbp at senescence. Conclusions: Imaging analysis of cell morphology is a useful tool for evaluating aging in cell cultures throughout the lifespan of MSCs. Our findings suggest that imaging analysis can reproducibly detect aging-related changes in cell morphology in MSC cultures. These findings suggest that cell morphology is still a supreme measure of cell quality and may be utilized to develop new noninvasive imaging-based methods to screen and quantitate aging in clinical-grade cell cultures.
Subject: Mesenchymal stromal cells
MSC
Aging
Senescence
Quality control
Morphology
Imaging
Cell manufacturing
Cell therapy
THERAPY POSITION STATEMENT
SENESCENT HUMAN-CELLS
STEM-CELLS
BONE-MARROW
INTERNATIONAL-SOCIETY
HUMAN FIBROBLASTS
ADIPOGENIC DIFFERENTIATION
OSTEOGENIC DIFFERENTIATION
REPLICATIVE SENESCENCE
TELOMERE LENGTH
3111 Biomedicine
1182 Biochemistry, cell and molecular biology
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