Embryoid body arrays: Parallel cryosectioning of spheroid/embryoid body samples for medium through-put analysis

Ross Ferguson, Vasanta Subramanian

Research output: Contribution to journalArticle

Abstract

Three dimensional (3D) culture of mammalian cells is emerging as a powerful new tool to understand organogenesis as well as serve as models for diseases with implications for therapeutic evaluation. 3D cultures are referred to variously as spheroid, organoids or embryoid bodies. While many methods exist for large scale production of embryoid bodies or other spheroid cell aggregates, either at controlled sizes using microwell/micropatterned plates or uncontrolled sizes in suspension dishes, very few protocols exist for medium throughput analysis of differentiation at the histological level. We have developed a method which allows for parallel processing, sectioning and analysis of multiple 3D samples (e.g. fixed at different time points, treated with different drugs/growth factors, generated from different cell lines etc.) by double-embedding blocks in a larger array format. Our protocol has few barriers for use and requires only materials commonly found in any lab currently using embedding materials for cryosectioning. Sectioning in parallel allows histological techniques (such as histochemistry, immunostaining or in situ hybridisation) to be performed simultaneously on many samples on a single slide. This reduces slide to slide variation as well as requiring less reagents, fewer consumables with lower time and labour requirements when compared to individually embedded samples.

LanguageEnglish
Pages125-130
Number of pages6
JournalStem Cell Research
Volume28
Early online date9 Feb 2018
DOIs
StatusPublished - 1 Apr 2018

Fingerprint

Cryoultramicrotomy
Embryoid Bodies
Organoids
Histological Techniques
Organogenesis
In Situ Hybridization
Intercellular Signaling Peptides and Proteins
Suspensions
Cell Culture Techniques
Cell Line
Pharmaceutical Preparations
Therapeutics

Keywords

  • Journal Article

Cite this

Embryoid body arrays : Parallel cryosectioning of spheroid/embryoid body samples for medium through-put analysis. / Ferguson, Ross; Subramanian, Vasanta.

In: Stem Cell Research, Vol. 28, 01.04.2018, p. 125-130.

Research output: Contribution to journalArticle

@article{bdfa045bb39d4d60bde4ae506a2b044c,
title = "Embryoid body arrays: Parallel cryosectioning of spheroid/embryoid body samples for medium through-put analysis",
abstract = "Three dimensional (3D) culture of mammalian cells is emerging as a powerful new tool to understand organogenesis as well as serve as models for diseases with implications for therapeutic evaluation. 3D cultures are referred to variously as spheroid, organoids or embryoid bodies. While many methods exist for large scale production of embryoid bodies or other spheroid cell aggregates, either at controlled sizes using microwell/micropatterned plates or uncontrolled sizes in suspension dishes, very few protocols exist for medium throughput analysis of differentiation at the histological level. We have developed a method which allows for parallel processing, sectioning and analysis of multiple 3D samples (e.g. fixed at different time points, treated with different drugs/growth factors, generated from different cell lines etc.) by double-embedding blocks in a larger array format. Our protocol has few barriers for use and requires only materials commonly found in any lab currently using embedding materials for cryosectioning. Sectioning in parallel allows histological techniques (such as histochemistry, immunostaining or in situ hybridisation) to be performed simultaneously on many samples on a single slide. This reduces slide to slide variation as well as requiring less reagents, fewer consumables with lower time and labour requirements when compared to individually embedded samples.",
keywords = "Journal Article",
author = "Ross Ferguson and Vasanta Subramanian",
note = "Copyright {\circledC} 2018. Published by Elsevier B.V.",
year = "2018",
month = "4",
day = "1",
doi = "10.1016/j.scr.2018.02.003",
language = "English",
volume = "28",
pages = "125--130",
journal = "Stem Cell Research",
issn = "1873-5061",
publisher = "Elsevier",

}

TY - JOUR

T1 - Embryoid body arrays

T2 - Stem Cell Research

AU - Ferguson, Ross

AU - Subramanian, Vasanta

N1 - Copyright © 2018. Published by Elsevier B.V.

PY - 2018/4/1

Y1 - 2018/4/1

N2 - Three dimensional (3D) culture of mammalian cells is emerging as a powerful new tool to understand organogenesis as well as serve as models for diseases with implications for therapeutic evaluation. 3D cultures are referred to variously as spheroid, organoids or embryoid bodies. While many methods exist for large scale production of embryoid bodies or other spheroid cell aggregates, either at controlled sizes using microwell/micropatterned plates or uncontrolled sizes in suspension dishes, very few protocols exist for medium throughput analysis of differentiation at the histological level. We have developed a method which allows for parallel processing, sectioning and analysis of multiple 3D samples (e.g. fixed at different time points, treated with different drugs/growth factors, generated from different cell lines etc.) by double-embedding blocks in a larger array format. Our protocol has few barriers for use and requires only materials commonly found in any lab currently using embedding materials for cryosectioning. Sectioning in parallel allows histological techniques (such as histochemistry, immunostaining or in situ hybridisation) to be performed simultaneously on many samples on a single slide. This reduces slide to slide variation as well as requiring less reagents, fewer consumables with lower time and labour requirements when compared to individually embedded samples.

AB - Three dimensional (3D) culture of mammalian cells is emerging as a powerful new tool to understand organogenesis as well as serve as models for diseases with implications for therapeutic evaluation. 3D cultures are referred to variously as spheroid, organoids or embryoid bodies. While many methods exist for large scale production of embryoid bodies or other spheroid cell aggregates, either at controlled sizes using microwell/micropatterned plates or uncontrolled sizes in suspension dishes, very few protocols exist for medium throughput analysis of differentiation at the histological level. We have developed a method which allows for parallel processing, sectioning and analysis of multiple 3D samples (e.g. fixed at different time points, treated with different drugs/growth factors, generated from different cell lines etc.) by double-embedding blocks in a larger array format. Our protocol has few barriers for use and requires only materials commonly found in any lab currently using embedding materials for cryosectioning. Sectioning in parallel allows histological techniques (such as histochemistry, immunostaining or in situ hybridisation) to be performed simultaneously on many samples on a single slide. This reduces slide to slide variation as well as requiring less reagents, fewer consumables with lower time and labour requirements when compared to individually embedded samples.

KW - Journal Article

U2 - 10.1016/j.scr.2018.02.003

DO - 10.1016/j.scr.2018.02.003

M3 - Article

VL - 28

SP - 125

EP - 130

JO - Stem Cell Research

JF - Stem Cell Research

SN - 1873-5061

ER -