Application of digital image analysis on histological images of a murine embryoid body model for monitoring endothelial differentiation
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Application of digital image analysis on histological images of a murine embryoid body model for monitoring endothelial differentiation. / Mousavi, Nabi; Raft, Marie Birkvig; Truelsen, Sarah Line Bring; Timmermans, Vera; Thastrup, Jacob; Heegaard, Steffen.
In: Pathology Research and Practice, Vol. 216, No. 11, 153225, 2020.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Application of digital image analysis on histological images of a murine embryoid body model for monitoring endothelial differentiation
AU - Mousavi, Nabi
AU - Raft, Marie Birkvig
AU - Truelsen, Sarah Line Bring
AU - Timmermans, Vera
AU - Thastrup, Jacob
AU - Heegaard, Steffen
PY - 2020
Y1 - 2020
N2 - The in vitro 3D model established from murine pluripotential stem cells (i.e., embryoid bodies (EBs)) is a dynamic model for endothelial differentiation. The aim of the present study was to investigate whether digital image analysis (DIA) can be applied on histological sections of EBs in order to quantify endothelial differentiation over time. The EBs were established in suspension cultures for 21 days in three independent replicate experiments. At day 4, 6, 9, 14, 18, and 21, the EBs were fixed in formaldehyde, embedded in paraffin and immunohistochemically (IHC) stained for CD31. The IHC-stained slides were digitally scanned and analysed using the Visiopharm® Quantitative Digital Pathology software Oncotopix™. The EBs developed CD31+ vascular-like structures during their differentiation. The quantitative DIA of the EBs showed that the log10 values of the relative CD31+ areas increased from −0.574 ± 0.470 (mean ± SD) at day 4 to 0.093 ± 0.688 (mean ± SD) at day 21 (p < 0.001). The approach presented in this study is a fast, quantitative and reproducible alternative method for an otherwise time-consuming and observer-dependent histological investigation. The future perspectives for such a system would be implementation of a modified version of the method on different 3D cultures and IHC markers.
AB - The in vitro 3D model established from murine pluripotential stem cells (i.e., embryoid bodies (EBs)) is a dynamic model for endothelial differentiation. The aim of the present study was to investigate whether digital image analysis (DIA) can be applied on histological sections of EBs in order to quantify endothelial differentiation over time. The EBs were established in suspension cultures for 21 days in three independent replicate experiments. At day 4, 6, 9, 14, 18, and 21, the EBs were fixed in formaldehyde, embedded in paraffin and immunohistochemically (IHC) stained for CD31. The IHC-stained slides were digitally scanned and analysed using the Visiopharm® Quantitative Digital Pathology software Oncotopix™. The EBs developed CD31+ vascular-like structures during their differentiation. The quantitative DIA of the EBs showed that the log10 values of the relative CD31+ areas increased from −0.574 ± 0.470 (mean ± SD) at day 4 to 0.093 ± 0.688 (mean ± SD) at day 21 (p < 0.001). The approach presented in this study is a fast, quantitative and reproducible alternative method for an otherwise time-consuming and observer-dependent histological investigation. The future perspectives for such a system would be implementation of a modified version of the method on different 3D cultures and IHC markers.
KW - 3D cultures
KW - CD31 differentiation
KW - Digital pathology
KW - Embryoid body
KW - Murine pluripotential stem cells
U2 - 10.1016/j.prp.2020.153225
DO - 10.1016/j.prp.2020.153225
M3 - Journal article
C2 - 32987302
AN - SCOPUS:85091575819
VL - 216
JO - Pathology, Research and Practice
JF - Pathology, Research and Practice
SN - 0344-0338
IS - 11
M1 - 153225
ER -
ID: 260990726