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

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 d after their collection, ten fresh samples were processed at these time intervals and evaluated. The cellular yield of most WJ samples was 1.1–5.0×10(6) cells at 21–30 d after processing. As culture time increased, cell counts decreased. Statistical analysis of mean cell counts showed a significant reduction after 21 d. 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 d 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.

Transplanted Umbilical Cord Mesenchymal Stem Cells Modify the In Vivo Microenvironment Enhancing Angiogenesis and Leading to Bone Regeneration

“…On the contrary, the classical expansion protocols for BM-MSCs, accepted for clinical use by the regulatory agencies [4], still employ FBS with FGF-2 as the medium additional supplement, which provides excellent results [37,38]. However, with regard to the present study, it should be noted that no major changes were observed in the phenotype characteristics and properties of BM-MSCs expanded in the presence of PL or FBS [34,36,39].…”

Fabrication and Characterization of Heparin/Collagen Sponge for in Vitro Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells into Hepatocytes

“…Wharton's jelly-derived mesenchymal stem cells (WJ-MSC) were isolated, as reported elsewhere (24), from the umbilical cords obtained from the infants delivered by cesarean section while informed consent was obtained from their parents. The umbilical cord vein was flashed with PBS, the arteries were removed, and the rest were cut into small pieces.…”

Transplantation of umbilical cord–derived mesenchymal stem cells as a novel strategy to protect the central nervous system: technical aspects, preclinical studies, and clinical perspectives

“…Recently, a new method has been proposed for the isolation of MSCs from Wharton's jelly by the direct delivery of enzymatic solutions into the tissue, similar to the method used for the enzymatic digestion of the pancreas for Langerhans islet isolation (50). To avoid the disadvantages of enzymatic digestion, which has been shown to potentially alter cell proliferation and function, some groups have developed UC explant culture approaches that have been shown to be simple, reproducible, and efficient (63)(64)(65)(66). However, considering the heterogeneity of the processes used for cell isolation and expansion, and in view of their use for clinical applications, some groups have tried to develop optimized and standardized methods based on the use of xeno-and serum-free culture media (52,67,68) or by taking advantage of the plastic-adherence properties of MSCs that allow cell expansion without UC dissection or enzymatic digestion (69).…”