Nora Khan

Abstract

While embryonic stem cells are well known to give rise to tissues comprising all three germ layers, only recently was it found that embryonic-like stem cells could be derived from the postnatal mice testis in culture. Embryonic-like stem cells from postnatal testes have shown that they can undertake most if not all, the functions of embryonic stem cells. Most recent reports have demonstrated that somatic cells can be induced to pluripotent stem cells, mostly with the addition of genes. However, the aim of this work is to explore the potential of testicular cells to become pluripotent, including expression of pluripotency-associated genes, and embryonic stem cells morphology, without the addition of genes. To accomplish our aim, we have sacrificed 15 CD-1 Swiss albino mice (8-10 weeks) and harvested cells from the testes using both mechanical and enzymatic digestion technique to generate single cell suspension. Isolated cells from the testes at day 1 along with embryonic-like stem cells at day 14 have been characterized via Reverse Transcription - Polymerase Chain Reaction (RT-PCR) using germ cells marker VASA, embryonic stem cells markers Nanog, cripto TDGF1, Esg1, and Thy1. Furthermore, Immunophenotyping was performed via Flow Cytometer using embryonic-like stem cells marker SOX2, CD 90 (Thy1), CD15 (SSEA-1), CD117 (c-kit), and CD133. Our results showed success in culturing cells harvested from the testes, these cells were reprogrammed and had the characteristics of embryonic stem cell. RT-PCR results confirmed the expression of VASA gene at day 1; these cells did not express embryonic stem cells markers (Nanog, cripto TDGF1, Esg1, and Thy1). However, embryonic-like stem cells at day 14 showed the expression of the previously mentioned embryonic stem cells markers, and did not express Vasa, which is a germ cell marker. Immunophenotyping results showed a noticed elevation in embryonic stem cells markers (SOX2, CD 90, CD15, CD117, and CD133) at day 14 compared to those at day 1. Here we report that embryonic-like stem cells can be derived from postnatal mice testis. However, further studies are required to assure the optimum value of embryonic-like stem cells that is dedifferentiated from mice testis before being used clinically in human.

Department

Biotechnology Program

Degree Name

MS in Biotechnology

Graduation Date

2-1-2017

Submission Date

January 2017

First Advisor

Amleh, Asma

Committee Member 1

Abou Aishah, Khaled

Committee Member 2

Abdellatif, Ahmed

Extent

54 p.

Document Type

Master's Thesis

Rights

The author retains all rights with regard to copyright. The author certifies that written permission from the owner(s) of third-party copyrighted matter included in the thesis, dissertation, paper, or record of study has been obtained. The author further certifies that IRB approval has been obtained for this thesis, or that IRB approval is not necessary for this thesis. Insofar as this thesis, dissertation, paper, or record of study is an educational record as defined in the Family Educational Rights and Privacy Act (FERPA) (20 USC 1232g), the author has granted consent to disclosure of it to anyone who requests a copy.

Institutional Review Board (IRB) Approval

Approval has been obtained for this item

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