Rapid Isolation of Single Cells from Mouse and Human Teeth

J 2021

Rapid Isolation of Single Cells from Mouse and Human Teeth

KŘIVÁNEK, Jan, Josef LAVICKÝ, Thibault BOUDERLIQUE and Igor ADAMEYKO

Basic information

Original name

Rapid Isolation of Single Cells from Mouse and Human Teeth

Authors

KŘIVÁNEK, Jan (203 Czech Republic, guarantor, belonging to the institution), Josef LAVICKÝ (203 Czech Republic, belonging to the institution), Thibault BOUDERLIQUE and Igor ADAMEYKO

Edition

Jove-Journal of Visualized Experiments, CAMBRIDGE, JOURNAL OF VISUALIZED EXPERIMENTS, 2021, 1940-087X

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 1.424

RIV identification code

RIV/00216224:14110/21:00124075

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.3791/63043

UT WoS

000715060500029

Keywords in English

Mouse and Human Teeth; Single Cells; Rapid Isolation

Abstract

V originále

Mouse and human teeth represent challenging organs for quick and efficient cell isolation for single-cell transcriptomic or other applications. The dental pulp tissue, rich in the extracellular matrix, requires a long and tedious dissociation process that is typically beyond the reasonable time for single-cell transcriptomics. For avoiding artificial changes in gene expression, the time elapsed from euthanizing an animal until the analysis of single cells needs to be minimized. This work presents a fast protocol enabling to obtain single-cell suspension from mouse and human teeth in an excellent quality suitable for scRNA-seq (single-cell RNA-sequencing). This protocol is based on accelerated tissue isolation steps, enzymatic digestion, and subsequent preparation of final single-cell suspension. This enables fast and gentle processing of tissues and allows using more animal or human samples for obtaining cell suspensions with high viability and minimal transcriptional changes. It is anticipated that this protocol might guide researchers interested in performing the scRNA-seq not only on the mouse or human teeth but also on other extracellular matrix-rich tissues, including cartilage, dense connective tissue, and dermis.

Links

MUNI/H/1615/2018, interní kód MU

Name: Understanding of tooth developmental trajectories as a novel approach for dental regeneration

Investor: Masaryk University, Individual High risk/high gain projects

MUNI/IGA/1532/2020, interní kód MU

Name: Investigation of the role of FoxD1+ stem cells during craniofacial development (Acronym: FoxD1 stem cells in craniofacial development)

Investor: Masaryk University

Citovat

KŘIVÁNEK, Jan, Josef LAVICKÝ, Thibault BOUDERLIQUE and Igor ADAMEYKO. Rapid Isolation of Single Cells from Mouse and Human Teeth. Jove-Journal of Visualized Experiments. CAMBRIDGE: JOURNAL OF VISUALIZED EXPERIMENTS, 2021, vol. 2021, No 176, p. 1-12. ISSN 1940-087X. Available from: https://dx.doi.org/10.3791/63043.

@article{1832355,
   author = {Křivánek, Jan and Lavický, Josef and Bouderlique, Thibault and Adameyko, Igor},
   article_location = {CAMBRIDGE},
   article_number = {176},
   doi = {http://dx.doi.org/10.3791/63043},
   keywords = {Mouse and Human Teeth; Single Cells; Rapid Isolation},
   language = {eng},
   issn = {1940-087X},
   journal = {Jove-Journal of Visualized Experiments},
   title = {Rapid Isolation of Single Cells from Mouse and Human Teeth},
   url = {https://www.jove.com/t/63043/rapid-isolation-of-single-cells-from-mouse-and-human-teeth},
   volume = {2021},
   year = {2021}
}

TY  - JOUR
ID  - 1832355
AU  - Křivánek, Jan - Lavický, Josef - Bouderlique, Thibault - Adameyko, Igor
PY  - 2021
TI  - Rapid Isolation of Single Cells from Mouse and Human Teeth
JF  - Jove-Journal of Visualized Experiments
VL  - 2021
IS  - 176
SP  - 1-12
EP  - 1-12
PB  - JOURNAL OF VISUALIZED EXPERIMENTS
SN  - 1940087X
KW  - Mouse and Human Teeth
KW  - Single Cells
KW  - Rapid Isolation
UR  - https://www.jove.com/t/63043/rapid-isolation-of-single-cells-from-mouse-and-human-teeth
N2  - Mouse and human teeth represent challenging organs for quick and efficient cell isolation for single-cell transcriptomic or other applications. The dental pulp tissue, rich in the extracellular matrix, requires a long and tedious dissociation process that is typically beyond the reasonable time for single-cell transcriptomics. For avoiding artificial changes in gene expression, the time elapsed from euthanizing an animal until the analysis of single cells needs to be minimized. This work presents a fast protocol enabling to obtain single-cell suspension from mouse and human teeth in an excellent quality suitable for scRNA-seq (single-cell RNA-sequencing). This protocol is based on accelerated tissue isolation steps, enzymatic digestion, and subsequent preparation of final single-cell suspension. This enables fast and gentle processing of tissues and allows using more animal or human samples for obtaining cell suspensions with high viability and minimal transcriptional changes. It is anticipated that this protocol might guide researchers interested in performing the scRNA-seq not only on the mouse or human teeth but also on other extracellular matrix-rich tissues, including cartilage, dense connective tissue, and dermis.
ER  -

KŘIVÁNEK, Jan, Josef LAVICKÝ, Thibault BOUDERLIQUE and Igor ADAMEYKO. Rapid Isolation of Single Cells from Mouse and Human Teeth. \textit{Jove-Journal of Visualized Experiments}. CAMBRIDGE: JOURNAL OF VISUALIZED EXPERIMENTS, 2021, vol.~2021, No~176, p.~1-12. ISSN~1940-087X. Available from: https://dx.doi.org/10.3791/63043.

Detailed Information on Publication Record