Abstract

 Background and Objectives

 The isolation of mesenchymal stem cells (MSCs) from the Wharton jelly (WJ) without degradation of cellular surface receptors and with the protection of cellular function, proliferation, and viability is important for research and clinical applications. In this study we have established a simple and rapid protocol without enzymatic treatment taking less time for MSCs from WJ to be isolated.

 Materials and Methods

 Human umbilical cords were collected after full-term deliveries and transported to the laboratory in sterile phosphate-buffered saline (PBS). After the removal of cords vessels, the WJ sectioned were transferred into PBS and put on a shaker for 2 hours. The cord segments were discarded and the suspension was centrifuged and cultured in DMEM-LG supplemented with 10% FBS for 5 days. The plastic adherent cells were investigated for surface markers. Adipogenic, osteogenic, and chondrogenic differentiations were performed to investigate the mesenchymal nature.

 Results

 After 5 days, purified populations of spindle-shape mesenchymal stem cell appeared and the cells were then expanded until they reached subconfluence. The cell population doubling time at third (31 ± 6h) and fourth (58 ± 15h ) passages was lower than other passages. The expanded cells were positive for CD 90, CD105, CD73, and CD44 , and negative for CD34/45, CD133, and HLA-DR surface markers. WJ-MSCs showed the potential of the multilineage cell differentiation into adipogenic, osteogenic, and chondrogenic phenotypes.

 Conclusions

 This study indicates that this non-enzymatic protocol can result in the efficient isolation of MSCs from Wharton jelly with less time taken. The expanded cells expressed characteristic markers and presented typical functional properties of MSCs such as the differentiation capacities.