Effectiveness of protocol for the isolation of Wharton's Jelly stem cells in large-scale applications

A. Petsa, Ph.D; S. Gargani, BSc; A. Felesakis, Bsc; N. Grigoriadis, BSc; I. Grigoriadis, Bsc

Biogenea-Cellgenea Ltd, Tsimiski 43, GR-54623, Thessaloniki, Greece

Corresponding Author: Dr. Anastasia Petsa, Ph.D.

Corresponding Author's Institution: Biogenea-Cellgenea Ltd.

Abstract:

The Wharton's Jelly (WJ) of the umbilical cord (UC) is an excellent source of mesenchymal stem cells (MSCs) with a range of potential therapeutic applications. The present study was conducted to demonstrate the efficiency of the protocols used by Biogenea-Cellgenea Ltd. for isolation and expansion of WJ MSCs from donors across Greece.

Umbilical cord samples were collected from 599 females following childbirth and processed for WJ

MSC isolation. Stem cells were expanded using DMEM-based media and cell counts and overall viability figures derived using Trypan blue exclusion. To investigate the application of isolation and expansion protocols on samples received 1, 2, 3, 4 and 5 days after their collection, 10 fresh samples were processed at these time intervals and evaluated.

The cellular yield of most WJ samples was 1.1-5.0 ×106 cells at 21-30 days after processing. As culture time increased, cell counts decreased. Statistical analysis of mean cell counts showed a significant reduction after 21 days. Finally, we demonstrate for the first time that it is possible to obtain satisfactory cell numbers from samples processed 1, 2, 3, 4 and even 5 days after collection.

We have derived favourable data on the protocols used at Biogenea-Cellgenea Ltd. to isolate and culture MSCs from the WJ. Protocol choice is crucial when handling large numbers of samples on a

daily basis and should be made to ensure the best possible outcome.

Keywords:

Wharton's Jelly, mesenchymal stem cells, isolation, expansion, population data