New Simple and Rapid Method for Purification of Mesenchymal Stem Cells from the Human Umbilical Cord Wharton Jelly

Abstract

We have developed a simple and rapid method for isolation of human umbilical cord matrix stem cells (hUCMS). The umbilical cord contains a virtual inexhaustible source of adult stem cells. We have substantially modified, simplified, and improved previously reported hUCMS isolation procedures in terms of either used enzyme type, or digestion time, and substantially enhanced the final product yield and purity. The isolated hUCMS were positive for CD90, CD117, and SCF, and negative for CD31 and CD45 surface markers. mRNA and related proteins (i.e., Sox2, Oct4a, Nanog, ABCG2, and c-Myc) that coincide with an uncommitted cell status also were detected. hUCMS express genes and proteins for CD90 and Nestin that are associated with mesenchymal stem cells, as well as other genes that specifically relate to different embryonic germ layers, namely, Vimentin, Sox7, Sox17, FoxA2, E-cadherin, and N-cadherin. hUCMS showed multilineage cell differentiation potential into adipogenic, osteogenic, and neural cell phenotypes, under the influence of lineage-specific, differentiation culture media. Moreover, the basal expression of endocrine cell markers makes these cells seemingly suitable for endocrine cell phenotype differentiation. Noteworthy, Activin A induced hUCMS to acquire definitive endoderm cell markers.

Effects of maternal age on the expression of mesenchymal stem cell markers in the components of human umbilical cord

“…The results of this study show that CD105-, CD29-positive and CD34-negative MSCs present in the major components of human umbilical cord (AM and WJ, UA and UV) and this finding is supported by some previous studies [1,4,5,[25][26][27]. It was an interesting observation that endothelial lining of both the UA and UV showed expression of the MSCs markers CD29 and CD105.…”

Human Mesenchymal Stroma/Stem Cells Exchange Membrane Proteins and Alter Functionality During Interaction with Different Tumor Cell Lines

“…The cDNA reactions were performed for 10 min/25°C, 1 h/37°C and stopped at 72°C for 10 min. As a template, 2.5 mL of cDNA was used with primers specific for CD73 (sense: 5¢-CGC AAC AAT GGC ACA ATT AC-3¢; antisense: 5¢-CTC GAC ACT TGG TGC AAA GA-3¢; amplification product 241 bp [27]), CD90 (sense: 5¢-GGA CTG AGA TCC CAG AAC CA-3¢; antisense: 5¢-ACG AAG GCT CTG GTC CAC TA-3¢; amplification product 124 bp [28]), and CD105 (sense: 5¢-TGT CTC ACT TCA TGC CTC CAG CT-3¢; antisense: 5¢-AGG CTG TCC ATG TTG AGG CAG T-3¢; amplification product 378 bp [29]). As a control, b-actin polymerase chain reaction (PCR) (sense: 5¢-CGG ATG TCC ACG TCA CAC T-3¢; antisense: 5¢-CCA CTG GCA TCG TGA TGG A-3¢; amplification product 427 bp [30]) was performed (all primers customized by Eurofins, MWG GmbH).…”

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

“…These protocols have been performed with or without the removal of cord blood vessels, after the dissection of the cord into small pieces or with the total length of the cord without mincing (62). 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).…”