MSC Cultivation from Umbilical Cord(UC) Tissue

As part of our regular QA test in May 2023, we have randomly chosen two cryopreserved umbilical cord tissue samples (stored Y2012 and Y2017) from our MVE storage tank. The objective is to check if the mesenchymal stem cells (MSCs) remain viable after being cryopreserved for 11 and 6 years respectively.

The selected cord tissues were thawed rapidly at 37°C while the accompanied cryoprotectant (or DMSO) was subsequently removed through a series of immersion rinsing. Once done, tissue pieces were excised from the larger segments and explanted in culture dishes with the appropriate reagent culture medium together with nutrients.

After 3 weeks, we clearly observed cells (In Figure 1:as indicated by green arrows) growing around tissue explants under the microscope. In general, human mesenchymal stem cells (hMSCs) are known to be spindle-shaped fibroblast-like (translucent and elongated) in appearance.

A look at the below photo images, Pointer A shows spindled shaped elongated translucent cells, which are the viable MSCs (similar to those shown with the green arrows in the 1st photo) growing from the tissue explants. These MSCs will continue to proliferate on the culture dish as a monolayer until it reaches 100% confluency. All Point B indicated are pieces of cryopreserved UC tissue explant. Given the correct conditions (CO2 level, temperature, medium), viable MSCs will migrate from the cryopreserved tissue and adhere to the culture dish where they begin to proliferate. Point to note is that those white dots present in all the photos are just some debris/unattached cells floating around and they are of no significant values.

From these QA tests, it shows that MSCs, which are multipotent stem cells, are still viable and can be isolated from umbilical cords tissues even after being cryopreserved for more than 10 years with CRYOLIFE. This is a testimony of CRYOLIFE’s long term quality storage capability for our clients’ precious umbilical cord and cord blood specimen.