Comparative analysis of mesenchymal stem cells derived from amniotic membrane, umbilical cord and chorionic plate under serum-free condition
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE118808 | GEO | 2019/01/23
REPOSITORIES: GEO
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Project description:BackgroundMesenchymal stem cells (MSCs) have emerged as a promising regenerative tool, owing mainly to their multi-differentiation potential and immunosuppressive capacity. When compared with MSCs classically derived from the adult bone marrow (BM), MSCs of neonatal origins exhibit superior proliferation ability, lower immunogenicity, and possible lower incorporated mutation; hence, they are considered as an alternative source for clinical use. Several researches have focused on the biological differences among some neonatal MSCs cultured in serum-containing medium (SCM). However, since it has been reported that MSCs possess different biological characteristics when cultured in serum-free medium (SFM), these comparative studies in SCM cannot exactly represent the results under the serum-free Good Manufacturing Practice (GMP) standard.MethodsHere, MSCs were isolated from three neonatal tissues, namely amniotic membrane (AM), umbilical cord (UC), and chorionic plate (CP), from the same donor, and their morphologies, immunophenotypes, trilineage differentiation potentials, global gene expression patterns, and proliferation abilities were systematically compared under chemical-defined SFM.ResultsOur study demonstrated that these three neonatal MSCs exhibited a similar morphology and immunophenotypic pattern but various mesodermal differentiation potentials under SFM: amniotic membrane-derived MSCs showed a higher rate for osteogenic differentiation; chorionic plate-derived MSCs presented better adipogenic induction efficiency; and all these three neonatal MSCs exhibited similar chondrogenic potential. Moreover, by the analysis of global gene expression patterns, we speculated a possible higher proliferation ability of CP-MSCs in SFM, and we subsequently validated this conjecture.ConclusionsCollectively, these results suggest that MSCs of different neonatal origins possess different biological features in SFM and thus may represent an optimal choice for different clinical applications.
| S-EPMC6330472 | biostudies-literature
Project description:Comparative analysis of mesenchymal stem cells derived from amniotic membrane, umbilical cord and chorionic plate under serum-free condition
| PRJNA486841 | ENA
Project description:Here, we use single cell transcriptomics to characterize a dynamic senescence process of in vitro expanded MSC products from adipose tissue (AD), bone marrow (BM), placenta membrane (PM) and umbilical cord (UC). We found that cultured MSCs underwent progressive cell aging. As compared to perinatal MSCs (derived from PM and UC), adult MSCs (derived from AD and BM) showed higher degree of senescence and impaired immunosuppressive function. Moreover, we identified the transcriptional network regulating the intra-population functional diversity, which provides us guidance to drive reprogramming of determined MSCs lineage with specific cellular functions for a particular therapeutic intent.
2023-06-19 | GSE200161 | GEO
Project description:BackgroundMesenchymal stem cells (MSCs) have been extensively explored as a promising therapeutic agent in the field of bone tissue engineering due to their osteogenic differentiation ability. In this study, the osteogenic differential ability and the effect of fibronectin and laminin on the osteogenic differentiation in four types of MSCs derived from placental tissue are compared to determine the ideal source for bone reconstruction tissue engineering.ResultsThe present study examines the osteogenic differentiation levels of four types of MSCs using alizarin red staining and quantifies the calcium levels and alkaline phosphatase (ALP) activity. In addition, this study examines the osteoblast differentiation protein markers osterix, collagen I, osteopontin, and osteocalcin using a Western blot assay. qPCR and EdU labeling assays were employed to identify the kinetics of osteogenic differentiation. Calcium deposit levels, ALP activity, and osteopontin and osteocalcin concentrations were determined to confirm the role of Extracellular matrix (ECM) components role on the osteogenic differentiation of MSCs. The data demonstrated that MSCs isolated from different layers of placenta had different potentials to differentiate into osteogenic cells. Importantly, AM-MSCs and UC-MSCs differentiated into the osteoblast stage more efficiently and quickly than CM-MSCs and DC-MSCs, which was associated with a decrease in their proliferation ability. Among the different types of MSCs, AM-MSCs and UC-MSCs had a higher osteogenic differentiation potential induced by fibronectin due to enhanced phosphorylation during the Akt and ERK pathways.ConclusionsTaken together, these results indicate that AM-MSCs and UC-MSCs possess a higher osteogenic potential, and fibronectin can robustly enhance the osteogenic potential of the Akt and ERK pathways.
| S-EPMC6371545 | biostudies-literature
Project description:Human umbilical cord matrix-derived mesenchymal stromal cells (UCM-MSC) are advantageous since can be easily obtained and display special interest as universal and feasible add-on therapy for myocardial infarction (MI). In this study, UCM-MSC from two umbilical cords, UC-A and UC-B, were transplanted in a murine MI model to investigate consistency and durability of the therapeutic benefits. Both cellular products supported sustained and long-term beneficial therapeutic effect. In vitro, the two cell products displayed similar ability to induce the formation of vessel-like structures and comparable transcriptome in normoxia and hypoxia, apart from expression differences in a small subset of genes associated with MHC Class I. These findings support that UCM-MSC are strong candidates to assist the treatment of MI whilst calling for the discussion on methodologies to characterize and select best performing UCM-MSC before clinical application.
2021-01-07 | E-MTAB-9978 | biostudies-arrayexpress
Project description:Mesenchymal stem cells (MSC) are multipotent cells which can be obtained from several adult and fetal tissues including human umbilical cord units. We have recently shown that umbilical cord tissue (UC) is richer in MSC than umbilical cord blood (UCB) but their origin and characteristics in blood as compared to the cord remains unknown. Here we compared, for the first time, the exonic protein-coding and intronic noncoding RNA (ncRNA) expression profiles of MSC from match-paired UC and UCB samples, harvested from the same donors, processed simultaneously and under the same culture conditions. The patterns of intronic ncRNA expression in MSC from UC and UCB paired units were highly similar, indicative of their common donor origin. The respective exonic protein-coding transcript expression profiles, however, were significantly different. Hierarchical clustering based on protein-coding expression similarities grouped MSC according to their tissue location rather than original donor. Genes related to systems development, osteogenesis and immune system were expressed at higher levels in UCB, whereas genes related to cell adhesion, morphogenesis, secretion, angiogenesis and neurogenesis were more expressed in UC cells. These molecular differences verified in tissue-specific MSC gene expression may reflect functional activities influenced by distinct niches and should be considered when developing clinical protocols involving MSC from different sources. In addition, these findings reinforce our previous suggestion on the importance of banking the whole umbilical cord unit for research or future therapeutic use.
2009-09-18 | GSE18134 | GEO
Project description:Background:Streptococcus pneumoniae colonize the human nasopharynx in the form of biofilms. The biofilms act as bacterial reservoirs and planktonic bacteria from these biofilms can migrate to other sterile anatomical sites to cause pneumonia, otitis media (OM), bacteremia and meningitis. Human amniotic membrane contains numerous growth factors and antimicrobial activity; however, these have not been studied in detail. In this study, we prepared amniotic membrane extract and chorionic membrane extract (AME/CME) and evaluated their antibacterial and antibiofilm activities against S. pneumoniae using an in vitro biofilm model and in vivo OM rat model. Materials and Methods: The AME/CME were prepared and protein was quantified using DCTM (detergent compatible) method. The minimum inhibitory concentrations were determined using broth dilution method, and the synergistic effect of AME/CME with Penicillin-streptomycin was detected checkerboard. The in vitro biofilm and in vivo colonization of S. pneumoniae were studied using microtiter plate assay and OM rat model, respectively. The AME/CME-treated biofilms were examined using scanning electron microscope and confocal microscopy. To examine the constituents of AME/CME, we determined the proteins and peptides of AME/CME using tandem mass tag-based quantitative mass spectrometry. Results: AME/CME treatment significantly (p < 0.05) inhibited S. pneumoniae growth in planktonic form and in biofilms. Combined application of AME/CME and Penicillin-streptomycin solution had a synergistic effect against S. pneumoniae. Biofilms grown with AME/CME were thin, scattered, and unorganized. AME/CME effectively eradicated pre-established pneumococci biofilms and has a bactericidal effect. AME treatment significantly (p < 0.05) reduced bacterial colonization in the rat middle ear. The proteomics analysis revealed that the AME/CME contains hydrolase, ribonuclease, protease, and other antimicrobial proteins and peptides. Conclusion: AME/CME inhibits S. pneumoniae growth in the planktonic and biofilm states via its antimicrobial proteins and peptides. AME/CME are non-cytotoxic, natural human product; therefore, they may be used alone or with antibiotics to treat S. pneumoniae infections.
| S-EPMC5641382 | biostudies-literature
Project description:Mesenchymal stem cells (MSC) are multipotent cells which can be obtained from several adult and fetal tissues including human umbilical cord units. We have recently shown that umbilical cord tissue (UC) is richer in MSC than umbilical cord blood (UCB) but their origin and characteristics in blood as compared to the cord remains unknown. Here we compared, for the first time, the exonic protein-coding and intronic noncoding RNA (ncRNA) expression profiles of MSC from match-paired UC and UCB samples, harvested from the same donors, processed simultaneously and under the same culture conditions. The patterns of intronic ncRNA expression in MSC from UC and UCB paired units were highly similar, indicative of their common donor origin. The respective exonic protein-coding transcript expression profiles, however, were significantly different. Hierarchical clustering based on protein-coding expression similarities grouped MSC according to their tissue location rather than original donor. Genes related to systems development, osteogenesis and immune system were expressed at higher levels in UCB, whereas genes related to cell adhesion, morphogenesis, secretion, angiogenesis and neurogenesis were more expressed in UC cells. These molecular differences verified in tissue-specific MSC gene expression may reflect functional activities influenced by distinct niches and should be considered when developing clinical protocols involving MSC from different sources. In addition, these findings reinforce our previous suggestion on the importance of banking the whole umbilical cord unit for research or future therapeutic use. Cy5- and Cy3-labeled cRNAs were obtained using 300 ng total RNA as template for amplification of poly(A) RNA by T7-RNA polymerase with the Agilent Low RNA Input Fluorescent Linear Amplification kit. The T7-polymerase amplified cRNA labeling approach advantageously replaces the reverse-transcriptase cDNA labeling used in early microarray experiments, because T7-RNA polymerase labeling of cRNA preserves the strand orientation of the original mRNA template. Reverse-transcriptase labeling can eventually generate a complementary cDNA second strand and cause artifactual labeling of a target with the opposite sense to that of the original message. Hybridization of 825 ng of Cy3- or Cy5-labeled RNA (dye swap technical replicate) from each UC sample with its paired UCB sample was performed with Agilent in situ Hybridization kit-plus, as recommended by the manufacturer, using a total of 4 intron-exon 44K expression oligoarrays. This array comprises a total of 13,699 exonic probes representing different protein-coding genes, along with custom-designed intronic probes for the antisense or sense strand. Slides were washed and processed according to the Agilent Two-Color Microarray-Based Gene Expression Analysis protocol (Version 5.5) and scanned on a GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA, USA). Fluorescence intensities were extracted using Feature Extraction (FE) software (version 9.0; Agilent). A gene was considered expressed if its probe intensity was significantly higher than the local background intensity, as calculated by the FE software. Then, the software applies local background subtraction and corrects for unequal dye incorporation using the default LOWESS (locally weighted linear regression) method.
2009-09-18 | E-GEOD-18134 | biostudies-arrayexpress
Project description:Based on transcriptome sequencing analysis, umbilical cord mesenchymal stem cells and placental mesenchymal stem cells share a total of 10529 common genes (called hpcCS consensus genes), which are umbilical cord-placental-consistent DEGs. The UC-MSC-specific region (referred to as the UC-specific gene) has most of the DEG (383 genes). The AM-MSC specific region (referred to as the AM specific gene) has 311 DEGs. The CM-MSC specific region (referred to as the CM specific gene) has 182 DEGs. The CV-MSC specific region (referred to as a CV-specific gene) has 169 DEGs. These results suggest that hMSCs from the umbilical cord and placenta have similar gene expression patterns, but have their own specific genes. The specific genes are not only involved in cell cycle, cell division, cell death, cell growth and development, transcriptional regulation, DNA repair, DNA. Replication plays an important role in chromosomal stability and is also involved in the movement of cellular or subcellular components, cellular communication and cell projection tissue, cytokine secretion and hormonal metabolism. The results of transcriptome sequencing analysis explain to some extent the differences in the biological characteristics of MSCs from different sources at the gene level.
2019-05-17 | GSE131355 | GEO
Project description:Acute Kidney Injury (AKI) remains a significant challenge with limited therapeutic interventions. This study explored the use of pulsed-focused ultrasound (pFUS) to prime the kidney, followed by mesenchymal stem cell (MSC) therapy to treat AKI. Our data showed that MSCs can mitigate some of the adverse effects of AKI, with these effects being greater with the use of umbilical cord (UC)-MSCs compared to bone marrow (BM)-MSCs. Next, we examined the effect of pFUS on the acutely injured kidney and found that compared to a single sonication, repeated sonication using pFUS at low acoustic doses (LpFUS) resulted in a substantial reduction in pro-inflammatory and immuno-modulatory cytokines. We then investigated the synergetic effect of repeated LpFUS followed by UC-MSC therapy; here, we found the therapeutic effects of UC-MSCs were enhanced resulting in a significant reduction in the blood urea nitrogen (BUN), improved body weight, and modulation of the kidney microenvironment with the expression of cytokines that created a more anti-inflammatory and regenerative milieu. Detailed transcriptomic analysis of acutely injured kidney tissue samples treated with LpFUS+UC-MSCs showed up-regulation of mitochondrial genes, specially PGC1α and DRP1, mitochondrial-associated proteins involved in the respiratory chain, nuclear genes encoding mitochondrial proteins, mitochondrial function, and specific heat shock proteins (i.e., HSP70 and HSP90). In conclusion, our results show that the therapeutic effect of UC-MSCs for treating AKI can be augmented by priming the kidney with repeated LpFUS before UC-MSC therapy. This innovative approach holds promise for advancing the treatment of AKI.
2024-04-30 | GSE262787 | GEO