Facilitated expansion of human embryonic stem cells by single-cell enzymatic dissociation
(2007) In Stem Cells 25(7). p.1690-1696- Abstract
- Traditionally, human embryonic stem cells (hESCs) are propagated by mechanical dissection or enzymatic dissociation into clusters of cells. To facilitate up-scaling and the use of hESC in various experimental manipulations, such as fluorescence-activated cell sorting, electroporation, and clonal selection, it is important to develop new, stable culture systems based on single-cell enzymatic propagation. Here, we show that hESCs, which were derived and passaged by mechanical dissection, can be rapidly adjusted to propagation by enzymatic dissociation to single cells. As an indication of the stability of this culture system, we demonstrate that hESCs can be maintained in an undifferentiated, pluripotent, and genetically normal state for up... (More)
- Traditionally, human embryonic stem cells (hESCs) are propagated by mechanical dissection or enzymatic dissociation into clusters of cells. To facilitate up-scaling and the use of hESC in various experimental manipulations, such as fluorescence-activated cell sorting, electroporation, and clonal selection, it is important to develop new, stable culture systems based on single-cell enzymatic propagation. Here, we show that hESCs, which were derived and passaged by mechanical dissection, can be rapidly adjusted to propagation by enzymatic dissociation to single cells. As an indication of the stability of this culture system, we demonstrate that hESCs can be maintained in an undifferentiated, pluripotent, and genetically normal state for up to 40 enzymatic passages. We also demonstrate that a recombinant trypsin preparation increases clonal survival compared with porcine trypsin. Finally, we show that human foreskin fibroblast feeders are superior to the commonly used mouse embryonic fibroblast feeders in terms of their ability to prevent spontaneous differentiation after single-cell passaging. Importantly, the culture system is widely applicable and should therefore be of general use to facilitate reliable large-scale cultivation of hESCs, as well as their use in various experimental manipulations. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/646084
- author
- Ellerström, Catharina LU ; Strehl, Raimund ; Noaksson, Karin ; Hyllner, Johan and Semb, Henrik LU
- organization
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- enzymatic, single cells, human embryonic stem cell, human feeders, dissociation, expansion
- in
- Stem Cells
- volume
- 25
- issue
- 7
- pages
- 1690 - 1696
- publisher
- Oxford University Press
- external identifiers
-
- wos:000247722100012
- scopus:34547158898
- pmid:17379766
- ISSN
- 1549-4918
- DOI
- 10.1634/stemcells.2006-0607
- language
- English
- LU publication?
- yes
- id
- 392be802-2212-459b-ac41-9237c3edcde0 (old id 646084)
- alternative location
- http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17379766&dopt=Abstract
- date added to LUP
- 2016-04-01 16:46:58
- date last changed
- 2023-02-23 03:13:12
@article{392be802-2212-459b-ac41-9237c3edcde0,
abstract = {{Traditionally, human embryonic stem cells (hESCs) are propagated by mechanical dissection or enzymatic dissociation into clusters of cells. To facilitate up-scaling and the use of hESC in various experimental manipulations, such as fluorescence-activated cell sorting, electroporation, and clonal selection, it is important to develop new, stable culture systems based on single-cell enzymatic propagation. Here, we show that hESCs, which were derived and passaged by mechanical dissection, can be rapidly adjusted to propagation by enzymatic dissociation to single cells. As an indication of the stability of this culture system, we demonstrate that hESCs can be maintained in an undifferentiated, pluripotent, and genetically normal state for up to 40 enzymatic passages. We also demonstrate that a recombinant trypsin preparation increases clonal survival compared with porcine trypsin. Finally, we show that human foreskin fibroblast feeders are superior to the commonly used mouse embryonic fibroblast feeders in terms of their ability to prevent spontaneous differentiation after single-cell passaging. Importantly, the culture system is widely applicable and should therefore be of general use to facilitate reliable large-scale cultivation of hESCs, as well as their use in various experimental manipulations.}},
author = {{Ellerström, Catharina and Strehl, Raimund and Noaksson, Karin and Hyllner, Johan and Semb, Henrik}},
issn = {{1549-4918}},
keywords = {{enzymatic; single cells; human embryonic stem cell; human feeders; dissociation; expansion}},
language = {{eng}},
number = {{7}},
pages = {{1690--1696}},
publisher = {{Oxford University Press}},
series = {{Stem Cells}},
title = {{Facilitated expansion of human embryonic stem cells by single-cell enzymatic dissociation}},
url = {{http://dx.doi.org/10.1634/stemcells.2006-0607}},
doi = {{10.1634/stemcells.2006-0607}},
volume = {{25}},
year = {{2007}},
}