Title

Alpha-1 Antitrypsin Increases Sox9 and Wnt3a Expression In Mesenchymal Stem Cells

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Abstract

Mesenchymal stem cells (MSCs) and the protein alpha-1 antitrypsin (AAT) have been used together to treat immune and inflammatory diseases. A previous experiment showed that AAT overexpression leads to an increase in SOX9 and Wnt3a in MSCs. This study examined whether AAT causes a change in the relative expression of SOX9 and Wnt3 over time. It was hypothesized that AAT will increase SOX9 and Wnt3a expression up to a certain time and then start decreasing the levels. After passaging human bone marrow mesenchymal stem cells (HBM-MSCs), the cells were transferred to four 12-well plates that were divided into collection times of 15, 30, 60, 180, and 360 minutes. The cells were treated with either 0.1 mg/ml of AAT or 0.5 mg/ml in 1 ml of media. The control cells were grown separately in 1 ml of regular media. After isolating mRNA and performing reverse transcription, qPCR was conducted to analyze gene expression in relation to the control GAPDH. The results showed that for the 0.5 mg/ml AAT treatment, the relative expression of SOX9 and Wnt3a gradually increased, peaked at 3 hours and then started to decrease. SOX9 and Wnt3a exhibited different trends for 0.1 mg/ml. AAT overexpressing mesenchymal stem cells exhibited higher levels of SOX9 and Wnt3a, with 0.5 mg/ml AAT causing greater changes. Future studies can fine-tune the dosage and timing of AAT to provide better treatment results.

Location

Furman Hall 107

Start Date

3-28-2020 2:30 PM

Presentation Format

Oral Only

Group Project

No

COinS
 
Mar 28th, 2:30 PM

Alpha-1 Antitrypsin Increases Sox9 and Wnt3a Expression In Mesenchymal Stem Cells

Furman Hall 107

Mesenchymal stem cells (MSCs) and the protein alpha-1 antitrypsin (AAT) have been used together to treat immune and inflammatory diseases. A previous experiment showed that AAT overexpression leads to an increase in SOX9 and Wnt3a in MSCs. This study examined whether AAT causes a change in the relative expression of SOX9 and Wnt3 over time. It was hypothesized that AAT will increase SOX9 and Wnt3a expression up to a certain time and then start decreasing the levels. After passaging human bone marrow mesenchymal stem cells (HBM-MSCs), the cells were transferred to four 12-well plates that were divided into collection times of 15, 30, 60, 180, and 360 minutes. The cells were treated with either 0.1 mg/ml of AAT or 0.5 mg/ml in 1 ml of media. The control cells were grown separately in 1 ml of regular media. After isolating mRNA and performing reverse transcription, qPCR was conducted to analyze gene expression in relation to the control GAPDH. The results showed that for the 0.5 mg/ml AAT treatment, the relative expression of SOX9 and Wnt3a gradually increased, peaked at 3 hours and then started to decrease. SOX9 and Wnt3a exhibited different trends for 0.1 mg/ml. AAT overexpressing mesenchymal stem cells exhibited higher levels of SOX9 and Wnt3a, with 0.5 mg/ml AAT causing greater changes. Future studies can fine-tune the dosage and timing of AAT to provide better treatment results.