Generation of mesenchymal stem cell from human umbilical cord tissue using a combination enzymatic and mechanical disassociation method

Abstract

MSCs (mesenchymal stem cells) promise a great potential for regenerative medicine due to their unique properties of self-renewal, high plasticity, modulation of immune response and the flexibility for genetic modification. Therefore, the increasing demand for cellular therapy necessitates a larger-scale production of MSC; however, the technical and ethical issues had put a halt on it. To date, studies have shown that MSC could be derived from human UC (umbilical cord), which is once considered as clinical waste. We have compared the two conventional methods which are classic enzymatic digestion and explant method with our newly tailored enzymatic-mechanical disassociation method to generate UC-MSC. The generated UC-MSCs from the methods above were characterized based on their immunophenotyping, early embryonic transcription factors expression and mesodermal differentiation ability. Our results show that enzymatic-mechanical disassociation method increase the initial nucleated cell yield greatly (approximately 160-fold) and maximized the successful rate of UC-MSC generation. Enzymatic-mechanical disassociation-derived UC-MSC exhibited fibroblastic morphology and surface markers expression of CD105, CD73, CD29, CD90 and MHC class I. Furthermore, these cells constitutively express early embryonic transcription factors (Nanog, Oct-4, Sox-2 and Rex-1), as confirmed by RT-PCR, indicating their multipotency and high self-renewal capacity. They are also capable of differentiating into osteoblasts and adipocytes when given an appropriate induction. The present study demonstrates a new and efficient approach in generating MSC from UC, hence serving as ideal alternative source of mesenchymal stem cell for clinical and research use.

Differentiation of Human Umbilical Cord Mesenchymal Stem Cells Into Insulin Producing Beta Cells for Diabetic Therapy.

“…It gave us an yield of 1.5*10 5 ± 0.5*10 5 cells/8cm of UC after 12 days of processing ( Figure 1). These numbers were relatively low when compared to reports from other groups [17,35] and may be due to the variation in the initial quantity of the cord tissue used.…”

Genetic association study of UCMA/GRP and OPTN genes (PDB6 locus) with Paget's disease of bone

“…Furthermore, Sox2 is also involved in the maintenance of self-renewal of the osteoblastic lineage [31]. Recently, it was shown that mesenchymal stem cells derived from human umbilical cord constitutively express SOX2 and are capable of differentiating into osteoblast as well as adipocytes, indicating the involvement of SOX2 in osteoblast differentiation [32].…”

Optimization of human umbilical cord mesenchymal stem cell isolation and culture methods

“…However, the tissue explant method has low separation efficiency, and the enzyme digestion method has many varying protocols, none of which are stable and efficient. In this study, we have compared the conventional tissue explant method (Ma et al 2005;Qiao et al 2008), the conventional enzyme digestion method (Wang et al 2004;Seshareddy et al 2008;Tong et al 2011) and a modified enzyme Fig. 4 Identification of cell antigen markers of the thirdgeneration of hUCMSCs isolated by the modified enzyme digestion method (9100).…”