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Article: Nutrient supplemented serum-free medium increases cardiomyogenesis efficiency of human pluripotent stem cells.

TitleNutrient supplemented serum-free medium increases cardiomyogenesis efficiency of human pluripotent stem cells.
Authors
Issue Date2013
PublisherBaishideng Publishing Group. The Journal's web site is located at http://www.wjgnet.com/1948-0210/index.htm
Citation
World Journal of Stem Cells, 2013, v. 5 n. 3, p. 86-97 How to Cite?
AbstractAIM: To development of an improved p38 MAPK inhibitor-based serum-free medium for embryoid body cardiomyocyte differentiation of human pluripotent stem cells. METHODS: Human embryonic stem cells (hESC) differentiated to cardiomyocytes (CM) using a p38 MAPK inhibitor (SB203580) based serum-free medium (SB media). Nutrient supplements known to increase cell viability were added to SB medium. The ability of these supplements to improve cardiomyogenesis was evaluated by measurements of cell viability, total cell count, and the expression of cardiac markers via flow cytometry. An improved medium containing Soy hydrolysate (HySoy) and bovine serum albumin (BSA) (SupSB media) was developed and tested on 2 additional cell lines (H1 and Siu-hiPSC). Characterization of the cardiomyocytes was done by immunohistochemistry, electrophysiology and quantitative real-time reverse transcription-polymerase chain reaction. RESULTS: hESC cell line, HES-3, differentiating in SB medium for 16 d resulted in a cardiomyocyte yield of 0.07 +/- 0.03 CM/hESC. A new medium (SupSB media) was developed with the addition of HySoy and BSA to SB medium. This medium resulted in 2.6 fold increase in cardiomyocyte yield (0.21 +/- 0.08 CM/hESC). The robustness of SupSB medium was further demonstrated using two additional pluripotent cell lines (H1, hESC and Siu1, hiPSC), showing a 15 and 9 fold increase in cardiomyocyte yield respectively. The age (passage number) of the pluripotent cells did not affect the cardiomyocyte yields. Embryoid body (EB) cardiomyocytes formed in SupSB medium expressed canonical cardiac markers (sarcomeric alpha-actinin, myosin heavy chain and troponin-T) and demonstrated all three major phenotypes: nodal-, atrial- and ventricular-like. Electrophysiological characteristics (maximum diastolic potentials and action potential durations) of cardiomyocytes derived from SB and SupSB media were similar. CONCLUSION: The nutrient supplementation (HySoy and BSA) leads to increase in cell viability, cell yield and cardiac marker expression during cardiomyocyte differentiation, translating to an overall increase in cardiomyocyte yield.
Persistent Identifierhttp://hdl.handle.net/10722/203106
ISSN
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorTing, Sen_US
dc.contributor.authorLecina, Men_US
dc.contributor.authorChan, YCen_US
dc.contributor.authorTse, HFen_US
dc.contributor.authorReuveny, Sen_US
dc.contributor.authorOh, SKWen_US
dc.date.accessioned2014-09-19T11:30:49Z-
dc.date.available2014-09-19T11:30:49Z-
dc.date.issued2013en_US
dc.identifier.citationWorld Journal of Stem Cells, 2013, v. 5 n. 3, p. 86-97en_US
dc.identifier.issn1948-0210-
dc.identifier.urihttp://hdl.handle.net/10722/203106-
dc.description.abstractAIM: To development of an improved p38 MAPK inhibitor-based serum-free medium for embryoid body cardiomyocyte differentiation of human pluripotent stem cells. METHODS: Human embryonic stem cells (hESC) differentiated to cardiomyocytes (CM) using a p38 MAPK inhibitor (SB203580) based serum-free medium (SB media). Nutrient supplements known to increase cell viability were added to SB medium. The ability of these supplements to improve cardiomyogenesis was evaluated by measurements of cell viability, total cell count, and the expression of cardiac markers via flow cytometry. An improved medium containing Soy hydrolysate (HySoy) and bovine serum albumin (BSA) (SupSB media) was developed and tested on 2 additional cell lines (H1 and Siu-hiPSC). Characterization of the cardiomyocytes was done by immunohistochemistry, electrophysiology and quantitative real-time reverse transcription-polymerase chain reaction. RESULTS: hESC cell line, HES-3, differentiating in SB medium for 16 d resulted in a cardiomyocyte yield of 0.07 +/- 0.03 CM/hESC. A new medium (SupSB media) was developed with the addition of HySoy and BSA to SB medium. This medium resulted in 2.6 fold increase in cardiomyocyte yield (0.21 +/- 0.08 CM/hESC). The robustness of SupSB medium was further demonstrated using two additional pluripotent cell lines (H1, hESC and Siu1, hiPSC), showing a 15 and 9 fold increase in cardiomyocyte yield respectively. The age (passage number) of the pluripotent cells did not affect the cardiomyocyte yields. Embryoid body (EB) cardiomyocytes formed in SupSB medium expressed canonical cardiac markers (sarcomeric alpha-actinin, myosin heavy chain and troponin-T) and demonstrated all three major phenotypes: nodal-, atrial- and ventricular-like. Electrophysiological characteristics (maximum diastolic potentials and action potential durations) of cardiomyocytes derived from SB and SupSB media were similar. CONCLUSION: The nutrient supplementation (HySoy and BSA) leads to increase in cell viability, cell yield and cardiac marker expression during cardiomyocyte differentiation, translating to an overall increase in cardiomyocyte yield.-
dc.languageengen_US
dc.publisherBaishideng Publishing Group. The Journal's web site is located at http://www.wjgnet.com/1948-0210/index.htm-
dc.relation.ispartofWorld Journal of Stem Cellsen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleNutrient supplemented serum-free medium increases cardiomyogenesis efficiency of human pluripotent stem cells.en_US
dc.typeArticleen_US
dc.identifier.emailChan, YC: yauchi@graduate.hku.hken_US
dc.identifier.emailTse, HF: hftse@hkucc.hku.hk-
dc.identifier.authorityChan, YC=rp01502en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.4252/wjsc.v5.i3.86-
dc.identifier.pmid23904910-
dc.identifier.pmcidPMC3723792-
dc.identifier.hkuros237961en_US
dc.identifier.volume5en_US
dc.identifier.issue3-
dc.identifier.spage86en_US
dc.identifier.epage97en_US
dc.publisher.placeUnited States-

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