当前位置： SCI文献检索 > Stem Cell Research & Therapy期刊下所有文献 > Automated image analysis detects aging in clinical-grade mesenchymal stromal cell cultures.

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

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 p16INK4a and p21Cip1/Waf1, β-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 μm2. At p5 the cells began to enlarge resulting in a 4.8-fold increase at p6-9 as compared to p1. The expression of p16INK4a and activity of β-galactosidase had a strong correlation with the increase in cell area, whereas the expression of p21Cip1/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.

journal_name

Stem Cell Res Ther

journal_title

Stem cell research & therapy

authors

Oja S,Komulainen P,Penttilä A,Nystedt J,Korhonen M

doi

10.1186/s13287-017-0740-x

subject

Has Abstract

pub_date

2018-01-10 00:00:00

pages

issue

issn

1757-6512

pii

10.1186/s13287-017-0740-x

journal_volume

pub_type

杂志文章

在线工具

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