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: Doungpunta J., Santhi A., Sathanawongs A., Jarujinda Y., Oranratnachai A.
Format: Article
Language:English
Published: 2014
Online Access:http://www.scopus.com/inward/record.url?eid=2-s2.0-67349201700&partnerID=40&md5=b6caf682c1972e31893517d9ff9f4963
http://www.ncbi.nlm.nih.gov/pubmed/19339041
http://cmuir.cmu.ac.th/handle/6653943832/2825
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-28252014-08-30T02:25:26Z Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors Doungpunta J. Santhi A. Sathanawongs A. Jarujinda Y. Oranratnachai A. 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. 2014-08-30T02:25:26Z 2014-08-30T02:25:26Z 2009 Article 0093691X 10.1016/j.theriogenology.2009.02.020 19339041 THGNB http://www.scopus.com/inward/record.url?eid=2-s2.0-67349201700&partnerID=40&md5=b6caf682c1972e31893517d9ff9f4963 http://www.ncbi.nlm.nih.gov/pubmed/19339041 http://cmuir.cmu.ac.th/handle/6653943832/2825 English
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
language English
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 Article
author Doungpunta J.
Santhi A.
Sathanawongs A.
Jarujinda Y.
Oranratnachai A.
spellingShingle Doungpunta J.
Santhi A.
Sathanawongs A.
Jarujinda Y.
Oranratnachai A.
Fivefold increase in derivation rates of mouse embryonic stem cells after supplementation of the media with multiple factors
author_facet Doungpunta J.
Santhi A.
Sathanawongs A.
Jarujinda Y.
Oranratnachai A.
author_sort Doungpunta J.
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 2014
url http://www.scopus.com/inward/record.url?eid=2-s2.0-67349201700&partnerID=40&md5=b6caf682c1972e31893517d9ff9f4963
http://www.ncbi.nlm.nih.gov/pubmed/19339041
http://cmuir.cmu.ac.th/handle/6653943832/2825
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