Umar Fuaad, Mimi Zulaikha
(2016)
Conditioned Medium From Bone Marrow-Derived Mesenchymal Stem Cells For Ex Vivo Expansion Of Cardiac Stem Cells.
Masters thesis, Universiti Sains Malaysia.
Abstract
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
Actions (login required)
 |
View Item |
![[img]](http://eprints.usm.my/style/images/fileicons/application_pdf.png)
