Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells

Sel induk mesenkima (MSC) daripada sumsum tulang merembes faktorfaktor parakrin yang mampu merangsang pengaktifan sel induk endogen jantung (CSC) dan memperbaiki kefungsian jantung. Faktor-faktor ini boleh dijana melalui pelaziman secara in vitro. Tesis ini bertujuan untuk mengoptimumkan kondisi...

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Main Author:	Umar Fuaad, Mimi Zulaikha
Format:	Thesis
Language:	English
Published:	2016
Subjects:	RK1-715 Dentistry
Online Access:	http://eprints.usm.my/32201/1/MIMI_ZULAIKHA_UMAR_FUAAD_24%28NN%29.pdf
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spelling	my-usm-ep.322012019-04-12T05:25:17Z Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells 2016-08 Umar Fuaad, Mimi Zulaikha RK1-715 Dentistry Sel induk mesenkima (MSC) daripada sumsum tulang merembes faktorfaktor parakrin yang mampu merangsang pengaktifan sel induk endogen jantung (CSC) dan memperbaiki kefungsian jantung. Faktor-faktor ini boleh dijana melalui pelaziman secara in vitro. Tesis ini bertujuan untuk mengoptimumkan kondisi pertumbuhan MSC dan perumusan medium yang sesuai untuk menjana medium terlazim (CdM) yang mempunyai ciri-ciri “cytoprotective” terhadap CSC. MSC dipencilkan daripada tibia dan femura mencit (C57BL/6N) yang berusia 3-5 minggu secara pembilasan sumsum tulang atau dengan menghancurkan tulang berkenaan dan dicirikan menggunakan sitometri aliran. Tempoh pre-pelaziman, dan kepadatan sel sebelum memulakan pelaziman (Fasa I), perumusan media, oksigen, tempoh pelaziman, kesan pengumpulan berulang, dan kepekatan pengolahan (Fasa II) dinilai dan dioptimum berdasarkan kesan CdM yang dijana terhadap pertumbuhan CSC secara in vitro. Seterusnya, CdM yang telah dioptimumkan itu diuji terhadap kadar migrasi CSC. CdM dipekatkan sebanyak 8 kali kepekatan (Fasa III) dan diuji terhadap pertumbuhan CSC, dibandingkan dengan CdM yang tidak dipekatkan. Data dianalisa menggunakan Analisis varians (ANOVA) dan Ujian-T. Bone marrow-derived mesenchymal stem cells (MSCs) have been shown to secrete paracrine factors which can stimulate activation of endogenous cardiac stem cells (CSCs) and ameliorate infarcted heart function. These factors can be harvested through conditioning of MSCs in vitro. This thesis aimed to optimise MSC growth conditions and medium formulation for generating conditioned medium (CdM) with CSC-cytoprotective properties. Murine MSCs were isolated from tibia and femur bones of 3-5 weeks old C57BL6/N mice using flushed marrow or crushed bone and was characterised by flow cytometry. Preconditioning time and seeding density before initiation (Phase I), medium formulation, oxygen, conditioning time, effects of repeated harvesting, and treatment concentration (Phase II) were assessed and optimised based on the effects of the produced CdM on CSCs survival in vitro. Then, the optimized CdM were tested on CSC migration. To reduce metabolic waste, CdM were concentrated 8 times (Phase III) and tested on CSC survival, and compared to the crude CdM. All data were analysed using ANOVA and t-test 2016-08 Thesis http://eprints.usm.my/32201/ http://eprints.usm.my/32201/1/MIMI_ZULAIKHA_UMAR_FUAAD_24%28NN%29.pdf application/pdf en public masters Universiti Sains Malaysia Institut Perubatan & Pergigian Termaju (Advanced Medical & Dental Institute)
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	RK1-715 Dentistry
spellingShingle	RK1-715 Dentistry Umar Fuaad, Mimi Zulaikha Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
description	Sel induk mesenkima (MSC) daripada sumsum tulang merembes faktorfaktor parakrin yang mampu merangsang pengaktifan sel induk endogen jantung (CSC) dan memperbaiki kefungsian jantung. Faktor-faktor ini boleh dijana melalui pelaziman secara in vitro. Tesis ini bertujuan untuk mengoptimumkan kondisi pertumbuhan MSC dan perumusan medium yang sesuai untuk menjana medium terlazim (CdM) yang mempunyai ciri-ciri “cytoprotective” terhadap CSC. MSC dipencilkan daripada tibia dan femura mencit (C57BL/6N) yang berusia 3-5 minggu secara pembilasan sumsum tulang atau dengan menghancurkan tulang berkenaan dan dicirikan menggunakan sitometri aliran. Tempoh pre-pelaziman, dan kepadatan sel sebelum memulakan pelaziman (Fasa I), perumusan media, oksigen, tempoh pelaziman, kesan pengumpulan berulang, dan kepekatan pengolahan (Fasa II) dinilai dan dioptimum berdasarkan kesan CdM yang dijana terhadap pertumbuhan CSC secara in vitro. Seterusnya, CdM yang telah dioptimumkan itu diuji terhadap kadar migrasi CSC. CdM dipekatkan sebanyak 8 kali kepekatan (Fasa III) dan diuji terhadap pertumbuhan CSC, dibandingkan dengan CdM yang tidak dipekatkan. Data dianalisa menggunakan Analisis varians (ANOVA) dan Ujian-T. Bone marrow-derived mesenchymal stem cells (MSCs) have been shown to secrete paracrine factors which can stimulate activation of endogenous cardiac stem cells (CSCs) and ameliorate infarcted heart function. These factors can be harvested through conditioning of MSCs in vitro. This thesis aimed to optimise MSC growth conditions and medium formulation for generating conditioned medium (CdM) with CSC-cytoprotective properties. Murine MSCs were isolated from tibia and femur bones of 3-5 weeks old C57BL6/N mice using flushed marrow or crushed bone and was characterised by flow cytometry. Preconditioning time and seeding density before initiation (Phase I), medium formulation, oxygen, conditioning time, effects of repeated harvesting, and treatment concentration (Phase II) were assessed and optimised based on the effects of the produced CdM on CSCs survival in vitro. Then, the optimized CdM were tested on CSC migration. To reduce metabolic waste, CdM were concentrated 8 times (Phase III) and tested on CSC survival, and compared to the crude CdM. All data were analysed using ANOVA and t-test
format	Thesis
qualification_level	Master's degree
author	Umar Fuaad, Mimi Zulaikha
author_facet	Umar Fuaad, Mimi Zulaikha
author_sort	Umar Fuaad, Mimi Zulaikha
title	Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
title_short	Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
title_full	Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
title_fullStr	Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
title_full_unstemmed	Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells
title_sort	conditioned medium from bone marrow-derived mesenchymal stem cells for ex vivo expansion of cardiac stem cells
granting_institution	Universiti Sains Malaysia
granting_department	Institut Perubatan & Pergigian Termaju (Advanced Medical & Dental Institute)
publishDate	2016
url	http://eprints.usm.my/32201/1/MIMI_ZULAIKHA_UMAR_FUAAD_24%28NN%29.pdf
_version_	1747820542693474304

Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells

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