ADSC Proliferation on Various SiO2 Membranes

Overview:

The goal of this experiment was to determine which membrane type adipose derived stem cells (ADSCs) would proliferate best on. The ADSCs grew over a  four day period and were imaged on days 1, 2, 3, and 4.  The cells were counted on each of these days to determine growth rates on each of the substrates and the data was then placed into graphs for comparison.

Methods:

The membranes the cells were grown on included: 0.5um low porosity, 0.5um high porosity, 3um low porosity, 3um high porosity, and nonporous, as well as tissue culture plastic. Each of the membranes were built in the CytoVu style and placed into 24-well plates and each had a sample size of three, the 0.5 LP and 0.5 HP had a sample size of 2 . For the tissue culture plastic (TCP) with and without GelTrex, the top gasket of a CytoVu was placed at the bottom of a 24-well plate. The silicon was exposed to ozone but the TCP was not.  After being constructed, all of the wells to be used in the experiment were exposed to one hour of UV light to be sterilized. Each of the membranes and one set of the TCP wells were covered in 1% GelTrex with PBS for 30 minutes, after that time the GelTrex was removed and the plates were left in the BSC to dry for an additional 30 minutes.  The cells were passaged at P4 and seeded onto each of the surfaces at 700 cells/well.  All of the cells were grown in ADSC standard proliferation media, this media is made of MesenPro RS+ 2% MesenPro Supplement+ 1% L-Glutamine+ 1% Pen-Strep. Images of each of the wells were taken at 24hrs, 48hrs, 72hrs, and 96hrs. Each of the four corners were imaged for every membrane in 10x, this was done so the entire membrane could be counted. Both porous and non-porous regions of the membranes were counted separately so movement of the cells could be observed.  For the TCP, the entire gasket area was imaged which was approximately 16 images. Four images were used from each time point around the approximate same area.

Data:

The graphs below show the comparison of ADSC proliferation on each of the surfaces.

Conclusion:

The data shows that each of the substrates has approximately the same proliferation rates with the non-porous SiO2 having the best proliferation rate and the TCP without geltrex and the 0.5LP having the lowest growth rates. There is some concern with the 0.5LP samples as one of the two samples exhibited behavior unlike all of the other cells on porous substrates. Using this information, the 0.5LP samples and the TCP samples will be redone.