CloneSeq - Single-cell clonal 3D culture and analysis protocol

This CloneSeq protocol combines clonal expansion inside 3D hydrogel spheres and droplet-based RNA sequencing to resolve the limited sensitivity of single-cell approaches. CloneSeq can reveal rare subpopulations and support cellular stemness. CloneSeq can be adapted to different biological systems to...

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Detalles Bibliográficos
Autores principales: Sun, Xue, Bavli, Danny, Kozulin, Chen, Motzik, Alex, Buxboim, Amnon, Ram, Oren
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8488600/
https://www.ncbi.nlm.nih.gov/pubmed/34632413
http://dx.doi.org/10.1016/j.xpro.2021.100794
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author Sun, Xue
Bavli, Danny
Kozulin, Chen
Motzik, Alex
Buxboim, Amnon
Ram, Oren
author_facet Sun, Xue
Bavli, Danny
Kozulin, Chen
Motzik, Alex
Buxboim, Amnon
Ram, Oren
author_sort Sun, Xue
collection PubMed
description This CloneSeq protocol combines clonal expansion inside 3D hydrogel spheres and droplet-based RNA sequencing to resolve the limited sensitivity of single-cell approaches. CloneSeq can reveal rare subpopulations and support cellular stemness. CloneSeq can be adapted to different biological systems to discover rare subpopulations by leveraging clonal enhanced sensitivity. Important considerations include the hydrogel composition, adaptation of 3D cultured clones to the inDrops system, and inherent adhesive properties of the cells. CloneSeq is only validated for cell lines so far. For complete details on the use and execution of this protocol, please refer to (Bavli et al., 2021).
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spelling pubmed-84886002021-10-08 CloneSeq - Single-cell clonal 3D culture and analysis protocol Sun, Xue Bavli, Danny Kozulin, Chen Motzik, Alex Buxboim, Amnon Ram, Oren STAR Protoc Protocol This CloneSeq protocol combines clonal expansion inside 3D hydrogel spheres and droplet-based RNA sequencing to resolve the limited sensitivity of single-cell approaches. CloneSeq can reveal rare subpopulations and support cellular stemness. CloneSeq can be adapted to different biological systems to discover rare subpopulations by leveraging clonal enhanced sensitivity. Important considerations include the hydrogel composition, adaptation of 3D cultured clones to the inDrops system, and inherent adhesive properties of the cells. CloneSeq is only validated for cell lines so far. For complete details on the use and execution of this protocol, please refer to (Bavli et al., 2021). Elsevier 2021-09-29 /pmc/articles/PMC8488600/ /pubmed/34632413 http://dx.doi.org/10.1016/j.xpro.2021.100794 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Sun, Xue
Bavli, Danny
Kozulin, Chen
Motzik, Alex
Buxboim, Amnon
Ram, Oren
CloneSeq - Single-cell clonal 3D culture and analysis protocol
title CloneSeq - Single-cell clonal 3D culture and analysis protocol
title_full CloneSeq - Single-cell clonal 3D culture and analysis protocol
title_fullStr CloneSeq - Single-cell clonal 3D culture and analysis protocol
title_full_unstemmed CloneSeq - Single-cell clonal 3D culture and analysis protocol
title_short CloneSeq - Single-cell clonal 3D culture and analysis protocol
title_sort cloneseq - single-cell clonal 3d culture and analysis protocol
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8488600/
https://www.ncbi.nlm.nih.gov/pubmed/34632413
http://dx.doi.org/10.1016/j.xpro.2021.100794
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