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Optimization of human mesenchymal stem cells isolation quality assessment from adipose tissue for clinical application

Puh, Špela (2016) MOBT01 20152
Degree Projects in Molecular Biology
Popular Abstract
Use of stem cells from fat tissue for clinical application

Stem cells isolated from fat tissue (Adipose Stem Cells, ASC) are desired therapeutic agents due to their good accessibility, high yield, ease of isolation, and their growth and differentiation capabilities. Under long-term in vitro culturing, cells lose their capabilities to grow and divide, and acquire an aging (senescent) morphology. Senescent cells are still metabolically active; however they can no longer divide, thus losing their clinical potential.

The aim of my project was to evaluate and compare fat stem cells (ASC) from different dog donors. Further, we wanted to assess whether there are any differences between cells isolated from young and old dogs, and between... (More)
Use of stem cells from fat tissue for clinical application

Stem cells isolated from fat tissue (Adipose Stem Cells, ASC) are desired therapeutic agents due to their good accessibility, high yield, ease of isolation, and their growth and differentiation capabilities. Under long-term in vitro culturing, cells lose their capabilities to grow and divide, and acquire an aging (senescent) morphology. Senescent cells are still metabolically active; however they can no longer divide, thus losing their clinical potential.

The aim of my project was to evaluate and compare fat stem cells (ASC) from different dog donors. Further, we wanted to assess whether there are any differences between cells isolated from young and old dogs, and between males and females. As gender-specific hormones and age are well known to have effect on cells abilities to divide and grow, we hypothesized that cells isolated from younger donors will prove to be better for clinical applicability. Our second aim was, to compare, if different culturing methods, that is, different ways cells are grown in vitro, have any effect on their growth characteristics.

To evaluate cells resistance to chemical exposure and senescence, cells were exposed to increasing concentrations of two chemical agents: mitoxantrone (Mxt) and trichostatin A (TSA) that are known to prevent cell division and further growth. We performed a cell viability assay, and we stained the cells with a beta-galactosidase staining solution, that specifically dies senescent cells blue.

In line with our expectations was that both Mxt and TSA negatively effected cell division and growth capabilities, and induced senescence. We showed that fat stem cells isolated from young dogs were of better quality for clinical application than ASC cells isolated from old dogs. Additionally, we observed variation between cells isolated form male and female dog donors, which was in favour to cells derived from male dogs. Our results also indicated that, due to different modes of action, TSA appears to be a better tool for evaluation of cells, before these cells could be used in therapy.

In terms of different cell culturing method, our results did not show any significant differences between different cell culturing methods.



Supervisor: prof. Tamara Lah Turnšek, PhD
Master Degree Project in Molecular Biology, 60 credits, 2016
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
author
Puh, Špela
supervisor
organization
course
MOBT01 20152
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
8891329
date added to LUP
2016-09-09 15:06:36
date last changed
2016-09-09 15:06:36
@misc{8891329,
  author       = {Puh, Špela},
  language     = {eng},
  note         = {Student Paper},
  title        = {Optimization of human mesenchymal stem cells isolation quality assessment from adipose tissue for clinical application},
  year         = {2016},
}