Effects of growth substrates on apoptosis and proliferation of human ovarian tissue cultured in vitro
(2019) MOBM02 20191Degree Projects in Molecular Biology
- Popular Abstract
- Fertility preservation techniques: A new hope
Cancer has become a widespread disease and a global concern affecting people of all age groups. Advances in cancer diagnosis and therapies have considerably increased the survival rates in younger patients. Chemotherapy and radiation therapy are the most common types of treatments; however, these treatments can have adverse side effects on fertility in women. This leads to the need of fertility preservation before the damaging treatments are started. Fertility can be preserved by cryopreservation of oocytes and embryos or even ovarian tissue, which can be used for fertility treatments when the patient later on wishes to have children. Ovarian tissue cryopreservation is the only fertility... (More) - Fertility preservation techniques: A new hope
Cancer has become a widespread disease and a global concern affecting people of all age groups. Advances in cancer diagnosis and therapies have considerably increased the survival rates in younger patients. Chemotherapy and radiation therapy are the most common types of treatments; however, these treatments can have adverse side effects on fertility in women. This leads to the need of fertility preservation before the damaging treatments are started. Fertility can be preserved by cryopreservation of oocytes and embryos or even ovarian tissue, which can be used for fertility treatments when the patient later on wishes to have children. Ovarian tissue cryopreservation is the only fertility preservation option for young (prepubertal) cancer patients. However, this technique cannot help all patients and more developments are needed. In particular in leukemias, transplantation of ovarian tissue could cause malignant cells to reoccur. For this patient group, growth and maturation of follicles in vitro would be a much-needed assisted reproductive technique (ART).
In vitro ovarian follicle culture has been developed since the 90s to become a novel assisted reproductive technology for those patients who cannot receive ovary tissue transplants. In vitro ovarian follicle culture involves extraction of ovarian tissue from the patients and stimulating maturation of the oocytes under in vitro conditions. Robust protocols for complete growth and maturation of human oocytes in vitro are still missing. An optimal in vitro model could benefit from growth substrates or biomaterials on which the ovarian tissue can be cultured. The substrate/ biomaterial must be able to mimic the architecture and dynamics of the tissue as under in vivo conditions. The aim of this thesis work was to examine naturally occurring components of ovarian extracellular matrix (ECM) as growth substrates for ovarian tissue in vitro.
Extracellular matrix (ECM) components have been widely used as biomaterial for various in vitro culture systems due to their fundamental roles in maintaining tissue homeostasis and structure. Collagen, laminins, fibronectin are some of the more popularly known ECM components. They are known to provide, support to the growing tissue and also aid in cell communication, migration, proliferation and survival. Studies in mouse and humans suggested the localization of several types of laminins in the follicular and stromal regions of the ovarian tissue. Based on these findings Laminin-221 (LN221) was selected as one of the growth substrates for this study. Laminin-521 (LN521) and Laminin-511 (LN511) which have previously been used for embryonic stem cell culture and follicle culture was also used as growth substrates. Matrigel®, which is a commercially available ECM mixture, has shown promising results so far but has several disadvantages like undefined composition, batch to batch variability and animal derived components. So, the aim of this study was to examine if the laminins can support ovarian tissue in culture as well as Matrigel®.
To evaluate the effects of the different laminins, apoptotic assay (TUNEL assay and caspase-3 immunostaining) was performed to determine cell death and proliferative assays (Ki-67 immunostaining) were performed to assess the survival nature of the tissue. The results suggested that tissue cultured on LN221 had lower amount of apoptosis when compared to other culture conditions. The Ki-67 assay suggested the presence of proliferative follicles especially in tissue cultured on LN521, LN511 and LN221. Previous histological analysis of follicles had suggested the presence of a large number of healthy follicles in the LN221 after the culture period. These findings collectively suggest that the LN221 is an effective growth substrate for in vitro culture of human ovarian tissue. Analysis of more samples and developing an optimal analysis method will enable better interpretation of the results. Future prospects could include combining the LN221 in a three-dimensional culture system which could better mimic the overall architecture necessary for growth and maturation of the follicles. Overall, research in this field will benefit fertility preservation techniques for young cancer patients who are at the risk of becoming infertile due to chemo and radiation therapy.
Master’s Degree Project in Molecular Biology 30 credits 2019
Department of Biology, Lund University
Advisor: Pauliina Damdimopoulou
Karolinska Institutet (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8997035
- author
- Prakash, Chandana Rao
- supervisor
- organization
- course
- MOBM02 20191
- year
- 2019
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 8997035
- date added to LUP
- 2019-10-24 14:50:09
- date last changed
- 2019-10-24 14:50:09
@misc{8997035,
author = {{Prakash, Chandana Rao}},
language = {{eng}},
note = {{Student Paper}},
title = {{Effects of growth substrates on apoptosis and proliferation of human ovarian tissue cultured in vitro}},
year = {{2019}},
}