---
_id: '13033'
abstract:
- lang: eng
text: Current methods for assessing cell proliferation in 3D scaffolds rely on changes
in metabolic activity or total DNA, however, direct quantification of cell number
in 3D scaffolds remains a challenge. To address this issue, we developed an unbiased
stereology approach that uses systematic-random sampling and thin focal-plane
optical sectioning of the scaffolds followed by estimation of total cell number
(StereoCount). This approach was validated against an indirect method for measuring
the total DNA (DNA content); and the Bürker counting chamber, the current reference
method for quantifying cell number. We assessed the total cell number for cell
seeding density (cells per unit volume) across four values and compared the methods
in terms of accuracy, ease-of-use and time demands. The accuracy of StereoCount
markedly outperformed the DNA content for cases with ~ 10,000 and ~ 125,000 cells/scaffold.
For cases with ~ 250,000 and ~ 375,000 cells/scaffold both StereoCount and DNA
content showed lower accuracy than the Bürker but did not differ from each other.
In terms of ease-of-use, there was a strong advantage for the StereoCount due
to output in terms of absolute cell numbers along with the possibility for an
overview of cell distribution and future use of automation for high throughput
analysis. Taking together, the StereoCount method is an efficient approach for
direct cell quantification in 3D collagen scaffolds. Its major benefit is that
automated StereoCount could accelerate research using 3D scaffolds focused on
drug discovery for a wide variety of human diseases.
acknowledgement: The study was supported by Project No. CZ.02.1.01/0.0/0.0/16_019/0000787
“Fighting INfectious Diseases”, awarded by the MEYS CR, financed from EFRR, by the
Cooperatio Program, research area DIAG and research area MED/DIAG, by the profiBONE
project (TO01000309) benefitting from a € (1.433.000) grant from Iceland, Liechtenstein
and Norway through the EEA Grants and the Technology Agency of the Czech Republic
and by a Grant (#1926990) to PRM and SRC Biosciences from the National Science Foundation
(U.S. Public Health Service). The authors acknowledge the invaluable assistance
provided by Iveta Paurova via her support in terms of the provision of laboratory
services.
article_number: '7959'
article_processing_charge: No
article_type: original
author:
- first_name: Anna
full_name: Zavadakova, Anna
last_name: Zavadakova
- first_name: Lucie
full_name: Vistejnova, Lucie
last_name: Vistejnova
- first_name: Tereza
full_name: Belinova, Tereza
id: 0bf89b6a-d28b-11eb-8bd6-f43768e4d368
last_name: Belinova
- first_name: Filip
full_name: Tichanek, Filip
last_name: Tichanek
- first_name: Dagmar
full_name: Bilikova, Dagmar
last_name: Bilikova
- first_name: Peter R.
full_name: Mouton, Peter R.
last_name: Mouton
citation:
ama: Zavadakova A, Vistejnova L, Belinova T, Tichanek F, Bilikova D, Mouton PR.
Novel stereological method for estimation of cell counts in 3D collagen scaffolds.
Scientific Reports. 2023;13(1). doi:10.1038/s41598-023-35162-z
apa: Zavadakova, A., Vistejnova, L., Belinova, T., Tichanek, F., Bilikova, D., &
Mouton, P. R. (2023). Novel stereological method for estimation of cell counts
in 3D collagen scaffolds. Scientific Reports. Springer Nature. https://doi.org/10.1038/s41598-023-35162-z
chicago: Zavadakova, Anna, Lucie Vistejnova, Tereza Belinova, Filip Tichanek, Dagmar
Bilikova, and Peter R. Mouton. “Novel Stereological Method for Estimation of Cell
Counts in 3D Collagen Scaffolds.” Scientific Reports. Springer Nature,
2023. https://doi.org/10.1038/s41598-023-35162-z.
ieee: A. Zavadakova, L. Vistejnova, T. Belinova, F. Tichanek, D. Bilikova, and P.
R. Mouton, “Novel stereological method for estimation of cell counts in 3D collagen
scaffolds,” Scientific Reports, vol. 13, no. 1. Springer Nature, 2023.
ista: Zavadakova A, Vistejnova L, Belinova T, Tichanek F, Bilikova D, Mouton PR.
2023. Novel stereological method for estimation of cell counts in 3D collagen
scaffolds. Scientific Reports. 13(1), 7959.
mla: Zavadakova, Anna, et al. “Novel Stereological Method for Estimation of Cell
Counts in 3D Collagen Scaffolds.” Scientific Reports, vol. 13, no. 1, 7959,
Springer Nature, 2023, doi:10.1038/s41598-023-35162-z.
short: A. Zavadakova, L. Vistejnova, T. Belinova, F. Tichanek, D. Bilikova, P.R.
Mouton, Scientific Reports 13 (2023).
date_created: 2023-05-19T11:12:25Z
date_published: 2023-05-17T00:00:00Z
date_updated: 2023-08-01T14:46:06Z
day: '17'
ddc:
- '570'
department:
- _id: Bio
doi: 10.1038/s41598-023-35162-z
external_id:
isi:
- '000995271600104'
file:
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date_created: 2023-05-22T07:57:37Z
date_updated: 2023-05-22T07:57:37Z
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intvolume: ' 13'
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keyword:
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language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '05'
oa: 1
oa_version: Published Version
publication: Scientific Reports
publication_identifier:
issn:
- 2045-2322
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
related_material:
link:
- relation: erratum
url: https://doi.org/10.1038/s41598-023-37265-z
scopus_import: '1'
status: public
title: Novel stereological method for estimation of cell counts in 3D collagen scaffolds
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 13
year: '2023'
...