Details
| Originalsprache | Englisch |
|---|---|
| Aufsatznummer | 1143 |
| Fachzeitschrift | Cells |
| Jahrgang | 11 |
| Ausgabenummer | 7 |
| Publikationsstatus | Veröffentlicht - 28 März 2022 |
Abstract
Organoids represent the cellular composition of natural tissue. So called colonoids, organoids derived from colon tissue, are a good model for understanding regeneration. However, next to the cellular composition, the surrounding matrix, the cell–cell interactions, and environmental factors have to be considered. This requires new approaches for the manipulation of a colonoid. Of key interest is the precise application of localized damage and the following cellular reaction. We have established multiphoton imaging in combination with femtosecond laser-based cellular nanosurgery in colonoids to ablate single cells in the colonoids’ crypts, the proliferative zones, and the differentiated zones. We observed that half of the colonoids recovered within six hours after manipulation. An invagination of the damaged cell and closing of the structure was observed. In about a third of the cases of targeted crypt damage, it caused a stop in crypt proliferation. In the majority of colonoids ablated in the crypt, the damage led to an increase in Wnt signalling, indicated via a fluorescent lentiviral biosensor. qRT-PCR analysis showed increased expression of various proliferation and Wnt-associated genes in response to damage. Our new model of probing colonoid regeneration paves the way to better understand organoid dynamics on a single cell level.
ASJC Scopus Sachgebiete
- Medizin (insg.)
- Allgemeine Medizin
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in: Cells, Jahrgang 11, Nr. 7, 1143, 28.03.2022.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Investigation of Colonic Regeneration via Precise Damage Application Using Femtosecond Laser-Based Nanosurgery
AU - Donath, Sören
AU - Angerstein, Leon
AU - Gentemann, Lara
AU - Müller, Dominik
AU - Seidler, Anna E.
AU - Jesinghaus, Christian
AU - Bleich, André
AU - Heisterkamp, Alexander
AU - Buettner, Manuela
AU - Kalies, Stefan
N1 - Funding Information: Funding: This study was funded by the REBIRTH Research Center for Translational Regenerative Medicine (ZN3440, State of Lower Saxony Ministry of Science and Culture (Nieders. Vorab)). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. M.B. and A.B. were funded by R2N, Federal State of Lower Saxony.
PY - 2022/3/28
Y1 - 2022/3/28
N2 - Organoids represent the cellular composition of natural tissue. So called colonoids, organoids derived from colon tissue, are a good model for understanding regeneration. However, next to the cellular composition, the surrounding matrix, the cell–cell interactions, and environmental factors have to be considered. This requires new approaches for the manipulation of a colonoid. Of key interest is the precise application of localized damage and the following cellular reaction. We have established multiphoton imaging in combination with femtosecond laser-based cellular nanosurgery in colonoids to ablate single cells in the colonoids’ crypts, the proliferative zones, and the differentiated zones. We observed that half of the colonoids recovered within six hours after manipulation. An invagination of the damaged cell and closing of the structure was observed. In about a third of the cases of targeted crypt damage, it caused a stop in crypt proliferation. In the majority of colonoids ablated in the crypt, the damage led to an increase in Wnt signalling, indicated via a fluorescent lentiviral biosensor. qRT-PCR analysis showed increased expression of various proliferation and Wnt-associated genes in response to damage. Our new model of probing colonoid regeneration paves the way to better understand organoid dynamics on a single cell level.
AB - Organoids represent the cellular composition of natural tissue. So called colonoids, organoids derived from colon tissue, are a good model for understanding regeneration. However, next to the cellular composition, the surrounding matrix, the cell–cell interactions, and environmental factors have to be considered. This requires new approaches for the manipulation of a colonoid. Of key interest is the precise application of localized damage and the following cellular reaction. We have established multiphoton imaging in combination with femtosecond laser-based cellular nanosurgery in colonoids to ablate single cells in the colonoids’ crypts, the proliferative zones, and the differentiated zones. We observed that half of the colonoids recovered within six hours after manipulation. An invagination of the damaged cell and closing of the structure was observed. In about a third of the cases of targeted crypt damage, it caused a stop in crypt proliferation. In the majority of colonoids ablated in the crypt, the damage led to an increase in Wnt signalling, indicated via a fluorescent lentiviral biosensor. qRT-PCR analysis showed increased expression of various proliferation and Wnt-associated genes in response to damage. Our new model of probing colonoid regeneration paves the way to better understand organoid dynamics on a single cell level.
KW - colonoid
KW - femtosecond laser
KW - lentiviral transduction
KW - nanosurgery
KW - organoid
UR - http://www.scopus.com/inward/record.url?scp=85127085009&partnerID=8YFLogxK
U2 - 10.3390/cells11071143
DO - 10.3390/cells11071143
M3 - Article
C2 - 35406708
AN - SCOPUS:85127085009
VL - 11
JO - Cells
JF - Cells
SN - 2073-4409
IS - 7
M1 - 1143
ER -