Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors

The objective of this study was to determine the ability of multiple-factor supplementation to augment derivation of mouse embryonic stem (mES) cells. Three factors, leukemia inhibitory factor (LIF), Parke-Davis 98059 (PD98059), and 6-bromoindirubin-3′-oxime (BIO), were added as supplements (individ...

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Main Authors: J. Doungpunta, A. Santhi, A. Sathanawongs, Y. Jarujinda, A. Oranratnachai
Format: Journal
Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/59266
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spelling th-cmuir.6653943832-592662018-09-10T03:25:56Z Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors J. Doungpunta A. Santhi A. Sathanawongs Y. Jarujinda A. Oranratnachai Agricultural and Biological Sciences Veterinary The objective of this study was to determine the ability of multiple-factor supplementation to augment derivation of mouse embryonic stem (mES) cells. Three factors, leukemia inhibitory factor (LIF), Parke-Davis 98059 (PD98059), and 6-bromoindirubin-3′-oxime (BIO), were added as supplements (individually or in a combination of all three) at two consecutive stages of culture; that is, from the start of blastocyst culture to the outgrowth stage, and from putting disaggregated outgrowth into culture medium to generation of primary mES colonies, respectively. The main outcome measure was the percentage of derivable mES cell lines, based on the number of blastocysts initially cultured. Three experiments demonstrated the following: (1) For the addition of individual single factor, only LIF yielded mES cell lines (6.2%), whereas a combination of all three factors resulted in the greatest number of mES cell lines (31.3%). (2) The advantages of a combination of multiple factors (LIF + PD98059 + BIO) were manifested only when they were used during the first stage of the culture and not during the second stage (31.6% vs. 6.2%, respectively). (3) The quality of the inner cell mass (ICM) outgrowth obtained from first-stage culture was studied. After alkaline phosphatase and Oct-4 staining, which documented pluripotency of the embryonic stem cells, outgrowths cultured in multiple factors (LIF + PD98059 + BIO) stained much stronger and in higher proportions than did those obtained after supplementation only with LIF (80% vs. 30%, respectively). © 2009 Elsevier Inc. All rights reserved. 2018-09-10T03:13:15Z 2018-09-10T03:13:15Z 2009-07-15 Journal 0093691X 2-s2.0-67349201700 10.1016/j.theriogenology.2009.02.020 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=67349201700&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/59266
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Agricultural and Biological Sciences
Veterinary
spellingShingle Agricultural and Biological Sciences
Veterinary
J. Doungpunta
A. Santhi
A. Sathanawongs
Y. Jarujinda
A. Oranratnachai
Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
description The objective of this study was to determine the ability of multiple-factor supplementation to augment derivation of mouse embryonic stem (mES) cells. Three factors, leukemia inhibitory factor (LIF), Parke-Davis 98059 (PD98059), and 6-bromoindirubin-3′-oxime (BIO), were added as supplements (individually or in a combination of all three) at two consecutive stages of culture; that is, from the start of blastocyst culture to the outgrowth stage, and from putting disaggregated outgrowth into culture medium to generation of primary mES colonies, respectively. The main outcome measure was the percentage of derivable mES cell lines, based on the number of blastocysts initially cultured. Three experiments demonstrated the following: (1) For the addition of individual single factor, only LIF yielded mES cell lines (6.2%), whereas a combination of all three factors resulted in the greatest number of mES cell lines (31.3%). (2) The advantages of a combination of multiple factors (LIF + PD98059 + BIO) were manifested only when they were used during the first stage of the culture and not during the second stage (31.6% vs. 6.2%, respectively). (3) The quality of the inner cell mass (ICM) outgrowth obtained from first-stage culture was studied. After alkaline phosphatase and Oct-4 staining, which documented pluripotency of the embryonic stem cells, outgrowths cultured in multiple factors (LIF + PD98059 + BIO) stained much stronger and in higher proportions than did those obtained after supplementation only with LIF (80% vs. 30%, respectively). © 2009 Elsevier Inc. All rights reserved.
format Journal
author J. Doungpunta
A. Santhi
A. Sathanawongs
Y. Jarujinda
A. Oranratnachai
author_facet J. Doungpunta
A. Santhi
A. Sathanawongs
Y. Jarujinda
A. Oranratnachai
author_sort J. Doungpunta
title Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
title_short Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
title_full Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
title_fullStr Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
title_full_unstemmed Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
title_sort fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=67349201700&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/59266
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