Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells

Human amniotic fluid (hAF) mesenchymal stem cells (MSCs) are commonly cultured in medium containing FBS. However, there are concerns about using animal serum in therapeutic applications due to the potential for immunogenic reactions and the risk of transmission of pathogens. For safety reasons, huma...

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Main Authors: Waleephan Tancharoen, Sirinda Aungsuchawan, Peraphan Pothacharoen, Kanokkan Bumroongkit, Chaniporn Puaninta, Nathaporn Pangjaidee, Suteera Narakornsak, Runchana Markmee, Tanongsak Laowanitwattana, Chawapon Thaojamnong
Format: Journal
Published: 2019
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/65369
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-653692019-08-05T04:40:15Z Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells Waleephan Tancharoen Sirinda Aungsuchawan Peraphan Pothacharoen Kanokkan Bumroongkit Chaniporn Puaninta Nathaporn Pangjaidee Suteera Narakornsak Runchana Markmee Tanongsak Laowanitwattana Chawapon Thaojamnong Biochemistry, Genetics and Molecular Biology Medicine Human amniotic fluid (hAF) mesenchymal stem cells (MSCs) are commonly cultured in medium containing FBS. However, there are concerns about using animal serum in therapeutic applications due to the potential for immunogenic reactions and the risk of transmission of pathogens. For safety reasons, human platelet lysate (hPL) has been suggested as a replacement for FBS because it appears to be a natural source of growth factors. In this present study, it was investigated whether FBS could be substituted with hPL in hAF‑MSCs culture without affecting their properties. Pooled hPL was generated by the freeze‑thaw method. The concentration of hPL was selected after evaluation by MTT assay. The hAF‑MSCs were cultured in FBS‑ or hPL‑supplemented conditions and shared a fibroblast‑like morphology. Cell proliferation assays showed that the growth characteristic of hAF‑MSCs cultured in 10% hPL‑supplemented media was similar to those cultured in 10% FBS‑supplemented media. The expression of MSC markers did not differ between the cells cultured in the different conditions. The endothelial differentiation potential was also investigated. Reverse transcription‑quantitative (RT‑q)PCR revealed that induced cells supplemented with hPL showed an increase level of endothelial specific gene expression compared to the FBS‑supplemented cells. Immunofluorescence analysis showed specific protein localization in both induced cell groups. Additionally, induced cells supplemented with hPL had the potential to form networks on Matrigel. This present study indicated that hPL could be used to culture and enhance the endothelial differentiation potential of hAF‑MSCs. 2019-08-05T04:32:15Z 2019-08-05T04:32:15Z 2019-06-01 Journal 17913004 2-s2.0-85066160234 10.3892/mmr.2019.10182 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85066160234&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/65369
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Biochemistry, Genetics and Molecular Biology
Medicine
spellingShingle Biochemistry, Genetics and Molecular Biology
Medicine
Waleephan Tancharoen
Sirinda Aungsuchawan
Peraphan Pothacharoen
Kanokkan Bumroongkit
Chaniporn Puaninta
Nathaporn Pangjaidee
Suteera Narakornsak
Runchana Markmee
Tanongsak Laowanitwattana
Chawapon Thaojamnong
Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
description Human amniotic fluid (hAF) mesenchymal stem cells (MSCs) are commonly cultured in medium containing FBS. However, there are concerns about using animal serum in therapeutic applications due to the potential for immunogenic reactions and the risk of transmission of pathogens. For safety reasons, human platelet lysate (hPL) has been suggested as a replacement for FBS because it appears to be a natural source of growth factors. In this present study, it was investigated whether FBS could be substituted with hPL in hAF‑MSCs culture without affecting their properties. Pooled hPL was generated by the freeze‑thaw method. The concentration of hPL was selected after evaluation by MTT assay. The hAF‑MSCs were cultured in FBS‑ or hPL‑supplemented conditions and shared a fibroblast‑like morphology. Cell proliferation assays showed that the growth characteristic of hAF‑MSCs cultured in 10% hPL‑supplemented media was similar to those cultured in 10% FBS‑supplemented media. The expression of MSC markers did not differ between the cells cultured in the different conditions. The endothelial differentiation potential was also investigated. Reverse transcription‑quantitative (RT‑q)PCR revealed that induced cells supplemented with hPL showed an increase level of endothelial specific gene expression compared to the FBS‑supplemented cells. Immunofluorescence analysis showed specific protein localization in both induced cell groups. Additionally, induced cells supplemented with hPL had the potential to form networks on Matrigel. This present study indicated that hPL could be used to culture and enhance the endothelial differentiation potential of hAF‑MSCs.
format Journal
author Waleephan Tancharoen
Sirinda Aungsuchawan
Peraphan Pothacharoen
Kanokkan Bumroongkit
Chaniporn Puaninta
Nathaporn Pangjaidee
Suteera Narakornsak
Runchana Markmee
Tanongsak Laowanitwattana
Chawapon Thaojamnong
author_facet Waleephan Tancharoen
Sirinda Aungsuchawan
Peraphan Pothacharoen
Kanokkan Bumroongkit
Chaniporn Puaninta
Nathaporn Pangjaidee
Suteera Narakornsak
Runchana Markmee
Tanongsak Laowanitwattana
Chawapon Thaojamnong
author_sort Waleephan Tancharoen
title Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
title_short Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
title_full Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
title_fullStr Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
title_full_unstemmed Human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
title_sort human platelet lysate as an alternative to fetal bovine serum for culture and endothelial differentiation of human amniotic fluid mesenchymal stem cells
publishDate 2019
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85066160234&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/65369
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