The Effect Of Media Supplements On Differentiation Of Human Adipose Derived Stem Cells Into Uroepithelial Cells

Sayaka Reed

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Jiro Nagatomi, Department of Bioengineering, Clemson University

Abstract

Recently, human Adipose Derived Stem Cells (hADSCs) have received attention as a potential cell source for organ regeneration, due to the ease of access as compared to bone marrow stem cells and the more controversial embryonic stem cells. Studies have demonstrated that ADSCs possess the multipotent capability of differentiating into several cell types including uroepithelial cells. They also possess immunosuppressant properties, making them easier to be accepted in organ transplants. Patients with bladder cancer have difficulty recovering from radiation treatment due to damage in the urinary epithelial lining. The damage leads to a lack of barrier function by the epithelium leading to cystitis. The purpose of this study is to test the hypothesis that cell culture media supplements can help differentiate hADSCs towards an uroepithelial cell lineage. The hADSCs were cultured in Keratinocyte Serum-Free Medium (KSFM) supplemented with All Trans-Retinoic Acid (ATRA), and their effects on specific cytokeratin and uroplakin uroepithelial markers were measured using RT-PCR and immunofluorescence at the mRNA and protein levels, respectively. The results indicate that there was an increase in the cytokeratins, specifically CK17 and CK20, and the uroplakins UPIb and UPII respectively. In addition, the control stem cell markers CD 90 and CD 105, which should have decreased, increased marginally. Although differentiation was successful, the results were incomparable to actual uroepithelial cells. The ultimate goal is to encapsulate the differentiated uroepithelial cells in hydrogels and apply them to damaged tissue to help in the recovery of the epithelial lining.

Start Date

4-11-2015 10:00 AM

End Date

4-11-2015 10:15 AM

COinS
 
Apr 11th, 10:00 AM Apr 11th, 10:15 AM

The Effect Of Media Supplements On Differentiation Of Human Adipose Derived Stem Cells Into Uroepithelial Cells

Recently, human Adipose Derived Stem Cells (hADSCs) have received attention as a potential cell source for organ regeneration, due to the ease of access as compared to bone marrow stem cells and the more controversial embryonic stem cells. Studies have demonstrated that ADSCs possess the multipotent capability of differentiating into several cell types including uroepithelial cells. They also possess immunosuppressant properties, making them easier to be accepted in organ transplants. Patients with bladder cancer have difficulty recovering from radiation treatment due to damage in the urinary epithelial lining. The damage leads to a lack of barrier function by the epithelium leading to cystitis. The purpose of this study is to test the hypothesis that cell culture media supplements can help differentiate hADSCs towards an uroepithelial cell lineage. The hADSCs were cultured in Keratinocyte Serum-Free Medium (KSFM) supplemented with All Trans-Retinoic Acid (ATRA), and their effects on specific cytokeratin and uroplakin uroepithelial markers were measured using RT-PCR and immunofluorescence at the mRNA and protein levels, respectively. The results indicate that there was an increase in the cytokeratins, specifically CK17 and CK20, and the uroplakins UPIb and UPII respectively. In addition, the control stem cell markers CD 90 and CD 105, which should have decreased, increased marginally. Although differentiation was successful, the results were incomparable to actual uroepithelial cells. The ultimate goal is to encapsulate the differentiated uroepithelial cells in hydrogels and apply them to damaged tissue to help in the recovery of the epithelial lining.