scholarly journals DAPT inhibits the chondrogenesis of human umbilical cord mesenchymal stem cells

2015 ◽  
Vol 10 (1) ◽  
Author(s):  
Xiaoke Hu ◽  
Jiawen Zhang ◽  
Xinxin Shao ◽  
Ermei Luo ◽  
Li Yu
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Notch Signaling ◽  
Umbilical Cord ◽  
Quantitative Polymerase Chain Reaction ◽  
Butyl Ester ◽  
Notch Pathway ◽  
Human Umbilical Cord ◽  
Notch 1

AbstractNotch signaling plays a key role in cell proliferation and differentiation, and is important in several biological processes, but its role in the chondrogenesis of human umbilical cord mesenchymal stem cells (UC-MSCs) is still unknown. N-[N-(3,5- difluorophenacetyl-L-alanyl)]-(S)-phenylglycinet-butyl ester (DAPT) is the inhibitor of Notch pathway. The aim of this study is to investgate the influence of DAPT on the chondrogenesis of UC-MSCs. In our study, UC-MSCs were isolated from human umbilical cord and their characteristics were identified. The UC-MSCs were induced to differentiate into chondrocytes in vitro and treated with 5 μM DAPT. Glycosaminoglycan (GAG) and collagen type II (COL-2A1) were analyzed qualitatively and quantitatively. The gene expression of Notch-1, Hes-1, GAG and COL-2A1 were analyzed by quantitative polymerase chain reaction (qPCR). The UC-MSCs separated from human umbilical cord, followed the characteristics of Mesenchymal Stem Cells (MSCs). The gene expression of Notch-1 and Hes-1 decreased after chondrogenic induction but the percentage in G1 period and the content of GAG and COL-2A1 increased. The expression of all tested Notch signaling and proliferation genes declined when 5 μM DAPT was added, also the content of GAG and COL-2A1 also decreased. Our study revealed that Notch signaling exists in UC-MSCs and it may remain the proliferative activity of UC-MSCs. Once the chondrogenesis begins, Notch signaling strength decline evidently. DAPT inhibits the chondrogenesis of UC-MSCs.

scholarly journals Programming of human umbilical cord mesenchymal stem cells in vitro to promote pancreatic gene expression

2013 ◽  
Vol 8 (3) ◽  
pp. 769-774 ◽  
Author(s):  
HONGWU WANG ◽  
YUPING YANG ◽  
GUYU HO ◽  
XIAOBO LIN ◽  
WEIZHAO WU ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Human Umbilical Cord

Stemness gene expression profile analysis in human umbilical cord mesenchymal stem cells

2012 ◽  
Vol 237 (6) ◽  
pp. 709-719 ◽  
Author(s):  
Ming-Yao Meng ◽  
Wei Pang ◽  
Li-Hong Jiang ◽  
Yun-Hong Liu ◽  
Chuan-Yu Wei ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Gene Expression Profile ◽  
Umbilical Cord ◽  
Expression Profile ◽  
Profile Analysis ◽  
Human Umbilical Cord ◽  
Expression Profile Analysis

scholarly journals Evaluation of NLR Family CARD Domain Containing 3 and NLR Family CARD Domain Containing 5 Gene Expression in Interferon Gamma–Treated Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Aida Nahumi ◽  
Leila Pirdel ◽  
Asadollah Asadi ◽  
Arash Abdolmaleki
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Interferon Gamma ◽  
Wharton’S Jelly ◽  
Human Umbilical Cord ◽  
Wharton's Jelly ◽  
Card Domain ◽  
Ifn Γ

Background: Human umbilical cord Wharton’s jelly has provided a new source for mesenchymal stem cells (MSCs). The highly proliferative capacity with low immunogenicity and multi-differentiation potential of its stem cells make them applicable for transplantation purposes. Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) play various roles in antigen presentation of pathogens and damaged cells to suppress and/or modulate inflammation. Objectives: In this study, the expression levels of NLR family CARD domain containing 3 (NLRC3) and NLRC5 genes were analyzed and compared in both untreated and interferon gamma (IFN-γ)–treated Wharton’s jelly-derived MSCs (WJ-MSCs). Methods: MSCs were isolated from human umbilical cord Wharton’s jelly using standard tissue culture. The expression of NLRC5 and NLRC3 genes was analyzed in IFN-γ–treated WJ-MSCs (24 hours after treatment) and untreated WJ-MSCs (as a control) using quantitative real-time polymerase chain reaction (PCR). Results: It was found that IFN-γ treatment mimicking an inflammation scenario led to a statistically significant increase of NLRC3 and NLRC5 gene expression compared to untreated WJ-MSCs (P ≤ 0.05). Conclusions: It seems that higher expression of NLRC3 and NLRC5 genes in treated WJ-MSCs may make them a proper candidate to be used as a source for cell therapy in inflammatory conditions.

scholarly journals Dodging COVID-19 infection: low expression and localization of ACE2 and TMPRSS2 in multiple donor-derived lines of human umbilical cord-derived mesenchymal stem cells

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Jonathan J. Hernandez ◽  
Doyle E. Beaty ◽  
Logan L. Fruhwirth ◽  
Ana P. Lopes Chaves ◽  
Neil H. Riordan
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Western Blot ◽  
Viral Infection ◽  
Umbilical Cord ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Umbilical Cord ◽  
Human Lung Tissue ◽  
Low Expression

Abstract Background Mesenchymal stem cells derived from human umbilical cord (hUC-MSCs) have immunomodulatory properties that are of interest to treat novel coronavirus disease 2019 (COVID-19). Leng et al. recently reported that hUC-MSCs derived from one donor negatively expressed Angiotensin-Converting Enzyme 2 (ACE2), a key protein for viral infection along with Transmembrane Serine Protease 2 (TMPRSS2). The purpose of this study was to quantify the expression of ACE2 and TMPRSS2 in hUC-MSCs lots derived from multiple donors using molecular-based techniques in order to demonstrate their inability to be a host to SARS-CoV-2. Methods Expression of ACE2 and TMPRSS2 was analyzed in 24 lots of hUC-MSCs derived from Wharton's jelly via quantitative polymerase chain reaction (qPCR), Western Blot, immunofluorescence and flow cytometry using 24 different donors. Results hUC-MSCs had significantly lower ACE2 (p = 0.002) and TMPRSS2 (p = 0.008) expression compared with human lung tissue homogenates in Western blot analyses. Little to no expression of ACE2 was observed in hUC-MSC by qPCR, and they were not observable with immunofluorescence in hUC-MSCs cell membranes. A negative ACE2 and TMPRSS2 population percentage of 95.3% ± 15.55 was obtained for hUC-MSCs via flow cytometry, with only 4.6% ACE2 and 29.5% TMPRSS2 observable positive populations. Conclusions We have demonstrated negative expression of ACE2 and low expression of TMPRSS2 in 24 lots of hUC-MSCs. This has crucial implications for the design of future therapeutic options for COVID-19, since hUC-MSCs would have the ability to “dodge” viral infection to exert their immunomodulatory effects.

Differential Expression of Osteogenic Differentiation in Human Umbilical Cord Wharton’s Jelly-Derived Mesenchymal Stem Cells Treated with Demineralized Bone

2008 ◽  
Vol 55-57 ◽  
pp. 697-700 ◽  
Author(s):  
D. Dhitiseith ◽  
S. Honsawek
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Wharton’S Jelly ◽  
Human Umbilical Cord ◽  
Activity Assay ◽  
Rt Pcr ◽  
Wharton's Jelly ◽  
Osteogenic Lineage

Mesenchymal stem cells are multipotential cells capable of differentiating into osteoblasts, chondrocytes, adipocytes, tenocytes, and myoblasts. Wharton’s jelly consists of stem cells that are a rich source of primitive multipotent mesenchymal cells. Demineralized bone matrix (DBM) has been widely utilized as a biomaterial to promote new bone formation. We isolate and characterize umbilical cord Wharton’s Jelly-derived mesenchymal stem (UCMS) cells derived from Wharton’s jelly and examine the biological activity of DBM in this cell line. Osteoblast differentiation of the UCMS cells was determined using alkaline phosphatase (ALP) activity assay. To examine differential gene expression during osteogenic differentiation, total RNA was isolated from UCMS cells in the absence or presence of DBM on day7 and analyzed using osteogenesis cDNA gene array. The selected genes were verified using reverse transcriptase-polymerase chain reaction (RT-PCR) analyses. Wharton’s jelly derived cells could differentiate along an osteogenic lineage after treatment of DBM. The ALP activity assay showed that human UCMS cells could differentiate into osteogenic lineage. Gene expression of human UCMS cells treated with DBM for 7 days was analyzed by using cDNA array and RT-PCR analyses. We found that expression of RUNX2 and SMAD2 was upregulated whereas SMAD7 expression was downregulated as confirmed by RT-PCR. UCMS cells from a Wharton’s jelly of human umbilical cord could express osteogenesis genes for treatment with DBM. Wharton’s jelly from umbilical cord is a new source of mesenchymal stem cells that are readily available for application to bone tissue engineering.

scholarly journals miR-1-Mediated Induction of Cardiogenesis in Mesenchymal Stem Cells via Downregulation of Hes-1

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Feng Huang ◽  
Liang Tang ◽  
Zhen-fei Fang ◽  
Xin-qun Hu ◽  
Jia-yi Pan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Notch Signaling ◽  
Embryonic Stem ◽  
Notch Pathway ◽  
Cardiac Gene Expression ◽  
Cardiac Gene ◽  
Cardiac Lineage ◽  
Infected Cells

MicroRNAs (miRNAs, miRs) have the potential to control stem cells fate decisions. The cardiac- and skeletal-muscle-specific miRNA, miR-1, can regulate embryonic stem cells differentiation to cardiac lineage by suppressing gene expression of alternative lineages. Accordingly, we hypothesized that overexpression of miR-1 may also promote cardiac gene expression in mesenchymal stem cells. Since Notch signaling could inhibit muscle differentiation, a process in contrast with the effect of miR-1, miR-1-mediated repression of Notch signaling may contribute to the observed effects of miR-1 in mesenchymal stem cells. Thus, mesenchymal stem cells were infected by lentiviral vectors carrying miR-1, and cells expressing miR-1 were selected. Alterations in Notch signaling and cardiomyocyte markers, Nkx2.5, GATA-4, cTnT, and CX43, were identified by Western blot in the infected cells on days 1, 7, and 14. Our study showed that the downstream target molecule of Notch pathway, Hes-1, was obviously decreased in mesenchymal stem cells modified with miR-1, and overexpression of miR-1 promotes the specific cardiac gene expression in the infected cells. Knockdown of Hes-1 leads to the same effects on cell lineage decisions. Our results indicated that miR-1 promotes the differentiation of MSCs into cardiac lineage in part due to negative regulation of Hes-1.

Mechanistic effect of human umbilical cord blood derived mesenchymal stem cells on the submandibular salivary gland in ovariectomized rats

2018 ◽  
Vol 96 (1) ◽  
pp. 57-67 ◽  
Author(s):  
Zienab A. Gouda ◽  
Mohamed E. Ali Khalifa ◽  
Sally M. Shalaby ◽  
Samia Hussein
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Submandibular Gland ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Ovariectomized Rats ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord

We performed this study to understand the effect of human umbilical cord blood derived mesenchymal stem cells (hUCB-MSCs) on the submandibular gland after bilateral ovariectomy. For this, 21 adult female rats were distributed equally among 3 groups: the sham-operated group (SHAM); the ovariectomized group (OVX); and the OVX group that received repeated intravenous injections of the hUCB-MSCs (OVX + hUCB-MSCs). We used reverse transcription – PCR to analyze for the gene expression of AQPs 3, 4, 5, and BMP-6. The cellular localization and expression of human CD105, human CD34, proliferating nuclear antigen (PCNA), single-stranded DNA (ss-DNA), caspase 3, AQP1, and α smooth muscle actin (α-SMA) were determined immunohistochemically. In the OVX group, a significant decrease in the gene expression of AQP3, AQP4, and BMP6, as well as the acinar area % was detected, while area % of granular convoluted tubules (GCTs) showed a significant increase. A significant decrease in area % staining positively for AQP1 and α-SMA was noted. An obvious improvement in the structure of the submandibular gland was demonstrated in the group injected with hUCB-MSCs, as well as a significant increase in the gene expression of AQP3, AQP4, and BMP6. The acinar and GCT area %, as well as the different measured markers, were relatively normal. This demonstrates that E2-deficiency induces structural changes to the submandibular gland. Moreover, a definite amelioration of the structure and function of the submandibular gland was detected after the administration of hUCB-MSCs.

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