scholarly journals Monocytes and pyrophosphate promote mesenchymal stem cell viability and early osteogenic differentiation

2022 ◽  
Vol 33 (1) ◽  
Author(s):  
Sara Svensson ◽  
Michael Palmer ◽  
Johan Svensson ◽  
Anna Johansson ◽  
Håkan Engqvist ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Culture Media ◽  
Conditioned Media ◽  
Human Mscs ◽  
Media Type ◽  
Inflammatory Monocytes ◽  
Ppar Γ

AbstractPyrophosphate-containing calcium phosphate implants promote osteoinduction and bone regeneration. The role of pyrophosphate for inflammatory cell-mesenchymal stem cell (MSC) cross-talk during osteogenesis is not known. In the present work, the effects of lipopolysaccharide (LPS) and pyrophosphate (PPi) on primary human monocytes and on osteogenic gene expression in human adipose-derived MSCs were evaluated in vitro, using conditioned media transfer as well as direct effect systems. Direct exposure to pyrophosphate increased nonadherent monocyte survival (by 120% without LPS and 235% with LPS) and MSC viability (LDH) (by 16–19% with and without LPS). Conditioned media from LPS-primed monocytes significantly upregulated osteogenic genes (ALP and RUNX2) and downregulated adipogenic (PPAR-γ) and chondrogenic (SOX9) genes in recipient MSCs. Moreover, the inclusion of PPi (250 μM) resulted in a 1.2- to 2-fold significant downregulation of SOX9 in the recipient MSCs, irrespective of LPS stimulation or culture media type. These results indicate that conditioned media from LPS-stimulated inflammatory monocytes potentiates the early MSCs commitment towards the osteogenic lineage and that direct pyrophosphate exposure to MSCs can promote their viability and reduce their chondrogenic gene expression. These results are the first to show that pyrophosphate can act as a survival factor for both human MSCs and primary monocytes and can influence the early MSC gene expression.

scholarly journals Canine Bone Marrow Mesenchymal Stem Cell Conditioned Media Affect Bacterial Growth, Biofilm-Associated Staphylococcus aureus and AHL-Dependent Quorum Sensing

2020 ◽  
Vol 8 (10) ◽  
pp. 1478 ◽  
Author(s):  
Dobroslava Bujňáková ◽  
Anna Čuvalová ◽  
Milan Čížek ◽  
Filip Humenik ◽  
Michel Salzet ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bone Marrow ◽  
Stem Cell ◽  
Quorum Sensing ◽  
Mesenchymal Stem Cell ◽  
Bacterial Growth ◽  
Conditioned Media ◽  
Amyloid Β Peptide

The present study investigated the in vitro antibacterial, antibiofilm and anti-Quorum Sensing (anti-QS) activities of canine bone marrow mesenchymal stem cell-conditioned media (cBM MSC CM) containing all secreted factors <30 K, using a disc diffusion test (DDT), spectrophotometric Crystal Violet Assay (SCVA) and Bioluminescence Assay (BA) with QS-reporter Escherichia coli JM109 pSB1142. The results show a sample-specific bacterial growth inhibition (zones varied between 7–30 mm), statistically significant modulation of biofilm-associated Staphylococcus aureus and Escherichia coli bioluminescence (0.391 ± 0.062 in the positive control to the lowest 0.150 ± 0.096 in the experimental group, cf. 11,714 ± 1362 to 7753 ± 700, given as average values of absorbance A550 ± SD versus average values of relative light units to growth RLU/A550 ± SD). The proteomic analysis performed in our previous experiment revealed the presence of several substances with documented antibacterial, antibiofilm and immunomodulatory properties (namely, apolipoprotein B and D; amyloid-β peptide; cathepsin B; protein S100-A4, galectin 3, CLEC3A, granulin, transferrin). This study highlights that cBM MSC CM may represent an important new approach to managing biofilm-associated and QS signal molecule-dependent bacterial infections. To the best of our knowledge, there is no previous documentation of canine BM MSC CM associated with in vitro antibiofilm and anti-QS activity.

scholarly journals Use of conditioned media (CM) and xeno-free serum substitute on human adipose-derived stem cells (ADSCs) differentiation into urothelial-like cells

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10890
Author(s):  
Ban Al- kurdi ◽  
Nidaa A. Ababneh ◽  
Nizar Abuharfeil ◽  
Saddam Al Demour ◽  
Abdalla S. Awidi
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Stem Cell ◽  
Urinary Tract ◽  
Culture Media ◽  
Urothelial Cell ◽  
Conditioned Media ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential ◽  
Expression Levels

Background Congenital abnormalities, cancers as well as injuries can cause irreversible damage to the urinary tract, which eventually requires tissue reconstruction. Smooth muscle cells, endothelial cells, and urothelial cells are the major cell types required for the reconstruction of lower urinary tract. Adult stem cells represent an accessible source of unlimited repertoire of untransformed cells. Aim Fetal bovine serum (FBS) is the most vital supplement in the culture media used for cellular proliferation and differentiation. However, due to the increasing interest in manufacturing xeno-free stem cell-based cellular products, optimizing the composition of the culture media and the serum-type used is of paramount importance. In this study, the effects of FBS and pooled human platelet (pHPL) lysate were assessed on the capacity of human adipose-derived stem cells (ADSCs) to differentiate into urothelial-like cells. Also, we aimed to compare the ability of both conditioned media (CM) and unconditioned urothelial cell media (UCM) to induce urothelial differentiation of ADCS in vitro. Methods ADSCs were isolated from human lipoaspirates and characterized by flow cytometry for their ability to express the most common mesenchymal stem cell (MSCs) markers. The differentiation potential was also assessed by differentiating them into osteogenic and adipogenic cell lineages. To evaluate the capacity of ADSCs to differentiate towards the urothelial-like lineage, cells were cultured with either CM or UCM, supplemented with either 5% pHPL, 2.5% pHPL or 10% FBS. After 14 days of induction, cells were utilized for gene expression and immunofluorescence analysis. Results ADSCs cultured in CM and supplemented with FBS exhibited the highest upregulation levels of the urothelial cell markers; cytokeratin-18 (CK-18), cytokeratin-19 (CK-19), and Uroplakin-2 (UPK-2), with a 6.7, 4.2- and a 2-folds increase in gene expression, respectively. Meanwhile, the use of CM supplemented with either 5% pHPL or 2.5% pHPL, and UCM supplemented with either 5% pHPL or 2.5% pHPL showed low expression levels of CK-18 and CK-19 and no upregulation of UPK-2 level was observed. In contrast, the use of UCM with FBS has increased the levels of CK-18 and CK-19, however to a lesser extent compared to CM. At the cellular level, CK-18 and UPK-2 were only detected in CM/FBS supplemented group. Growth factor analysis revealed an increase in the expression levels of EGF, VEGF and PDGF in all of the differentiated groups. Conclusion Efficient ADSCs urothelial differentiation is dependent on the use of conditioned media. The presence of high concentrations of proliferation-inducing growth factors present in the pHPL reduces the efficiency of ADSCs differentiation towards the urothelial lineage. Additionally, the increase in EGF, VEGF and PDGF during the differentiation implicates them in the mechanism of urothelial cell differentiation.

scholarly journals The The Influence of Wharton Jelly Mesenchymal Stem Cell toward Matrix Metalloproteinase-13 and RELA Synoviocyte Gene Expression on Osteoarthritis

2019 ◽  
Vol 7 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Vivi Sofia ◽  
Ellyza Nasrul ◽  
Menkher Manjas ◽  
Gusti Revilla
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Inflammatory Responses ◽  
Group Iv ◽  
Group Vi ◽  
Group V ◽  
Group Iii ◽  
Group I

BACKGROUND: Therapy for osteoarthritis (OA) with satisfactory results has not been found to date. In OA pathogenesis, RELA gene involved in cartilage degradation and MMP-13 in degrade cartilage, as a member family of NF-ĸβ genes, RELA serves to modulate inflammatory responses and activates pro-inflammatory cytokines. AIM: This study aims to identify the influence of Wharton Jelly Mesenchymal Stem Cell (MSC-WJ) on MMP-13 and RELA expression gene in synoviocyte by in vitro. MATERIAL AND METHODS: This research is pure experimental research. The sample used derived from synovial tissue of OA patients who underwent Total Knee Replacement (TKR) surgery. This study was divided into six groups treated with 4 replications. Group I and II (control groups) were synoviocyte of OA incubated for 24 and 48 hours, respectively. Group III and IV were MSC-WJ incubated for 24 and 48 hours, respectively. Group V and VI were Synoviocyte-MSC-WJ co-culture group incubated for 24 and 48 hours, respectively. Identification of MMP-13 and RELA gene expression in each group was performed by using qPCR. RESULT: The results showed that MSC-WJ reduced MMP-13 gene expression after co-culture for 24 and 48 hours in OA synoviocyte. The highest gene expression of MMP-13 was in Group I and II (1.00 ng/μl), followed by Group III (0.41 ng/μl), Group IV (0.24 ng/μl), Group V (0.13 ng/μl), and Group VI (0.04 ng/μl). MSC-WJ administration also decreased RELA gene expression. The highest gene expression of RELA gene was in Group I and II (1.00 ng/μl), Group V (0.67 ng/μl), Group III (0.58 ng/μl), Group IV (0.16 ng/μl), and Group VI (0.16 ng/μl). CONCLUSION: This study concluded that MSC-WJ in OA synoviocyte significantly reduced the expression of MMP-13 and RELA gene (p <0.05).

scholarly journals Comparison of Anti-Oxidative Effect of Human Adipose- and Amniotic Membrane-Derived Mesenchymal Stem Cell Conditioned Medium on Mouse Preimplantation Embryo Development

Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 268
Author(s):  
Kihae Ra ◽  
Hyun Ju Oh ◽  
Eun Young Kim ◽  
Sung Keun Kang ◽  
Jeong Chan Ra ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Amniotic Membrane ◽  
Preimplantation Embryo ◽  
Preimplantation Embryo Development ◽  
Antioxidant Gene ◽  
Mouse Preimplantation Embryo ◽  
Antioxidant Gene Expression

Oxidative stress is a major cause of damage to the quantity and quality of embryos produced in vitro. Antioxidants are usually supplemented to protect embryos from the suboptimal in vitro culture (IVC) environment. Amniotic membrane-derived mesenchymal stem cells (AMSC) have emerged as a promising regenerative therapy, and their paracrine factors with anti-oxidative effects are present in AMSC conditioned medium (CM). We examined the anti-oxidative potential of human AMSC-CM treatment during IVC on mouse preimplantation embryo development and antioxidant gene expression in the forkhead box O (FoxO) pathway. AMSC-CM (10%) was optimal for overall preimplantation embryo developmental processes and upregulated the expression of FoxOs and their downstream antioxidants in blastocysts (BL). Subsequently, compared to adipose-derived mesenchymal stem cell (ASC)-CM, AMSC-CM enhanced antioxidant gene expression and intracellular GSH levels in the BL. Total antioxidant capacity and SOD activity were greater in AMSC-CM than in ASC-CM. Furthermore, SOD and catalase were more active in culture medium supplemented with AMSC-CM than in ASC-CM. Lastly, the anti-apoptotic effect of AMSC-CM was observed with the regulation of apoptosis-related genes and mitochondrial membrane potential in BL. In conclusion, the present study established AMSC-CM treatment at an optimal concentration as a novel antioxidant intervention for assisted reproduction.

scholarly journals The Analysis of the Relationship between RELA Gene Expression and MMP-13 Gene Expression in Synoviocyte Cells after Mesenchymal Stem Cell Wharton Jelly

2019 ◽  
Vol 7 (4) ◽  
pp. 543-548 ◽  
Author(s):  
Vivi Sofia ◽  
Ellyza Nasrul ◽  
Menkher Manjas ◽  
Gusti Revilla
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cartilage Damage ◽  
Cartilage Degradation ◽  
Cell Group ◽  
Control Group ◽  
Group I ◽  
The Relationship

BACKGROUND: Therapy that can cure osteoarthritis with satisfactory results has not been found to date. In the pathogenesis of osteoarthritis, the genes involved in cartilage degradation include the RELA gene which plays an important role in modulating the occurrence of cartilage damage, which involves activation of pro-inflammatory cytokines. One of the cytokines involved in the cartilage degradation process is Matrix Metalloproteinase (MMP) -13 which is also modulated by NFĸβ. AIM: This study aims to look at the expression of the RELA gene and expression of the MMP-13 gene and analyse the relationship of RELA gene expression with MMP-13 gene expression after administration of Mesenchymal Stem Cell Wharton Jelly in synoviocytes in vitro. MATERIAL AND METHODS: This research is pure experimental research. The samples used derived from synovial tissue in osteoarthritis patients who underwent surgery for Total Knee Replacement (TKR). This study was divided into 6 treatment groups with 4 replications. Group I was the synoviocyte OA cell control group which was incubated 24 hours, group II was control of synoviocyte OA cell which was incubated 48 hours, group III was a group of Mesenchymal Stem Cell Wharton Jelly (MSC-WJ) which was incubated 24 hours, group IV was a Mesenchymal Stem Cell Wharton Jelly (MSC-WJ) cell group incubated 48 hours, group V was the co-culture group of synoviocyte-MSC-WJ cells incubated 24 hours and group VI was the co-culture of synoviocyte-MSC-WJ cells which were incubated 48 hours. Observation of MMP-13 gene expression and RELA gene in each group was carried out using qPCR. RESULT: The results showed that the analysis of the relationship between RELA gene expression and MMP-13 gene expression in osteoarthritis synoviocytes cells after Mesenchymal Stem Cell Wharton Jelly as big as (r = 0.662). CONCLUSION: The conclusion of this study is there was a strong correlation between RELA gene expression and MMP-13 gene expression in osteoarthritis synoviocytes after Mesenchymal Stem Cell Wharton Jelly (r = 0.662).

scholarly journals Investigation of Stem Cell Applications on In Vitro Fertilization in Rats

2021 ◽  
Vol 72 (3) ◽  
pp. 3067
Author(s):  
A GUMURDU ◽  
S OZTURK ◽  
I AYDEMIR ◽  
MI TUGLU
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
In Vitro Fertilization ◽  
Mesenchymal Stem Cell ◽  
Conditioned Media ◽  
Female Rats ◽  
Favourable Effect ◽  
Vitro Fertilization ◽  
Male And Female Rats

We aimed to search the effects of bone marrow-derived mesenchymal stem cell-conditioned media on in vitro fertilization by investigation of lifetime of germ cells cleavage, degeneration rates and embryo quality. For this purpose, firstly MSCs were isolated from femurs and tibias of the rat, and cells were cultured until the fourth passage. Sperm and oocytes were collected from male and female rats. Oocytes were added in Human Tubal Fluid Media (HTFM), Single Step Media (SSM), Alpha-MEM Media (AMM) and Bone Marrow-Derived Mesenchymal Stem Cell-Conditioned Media (CM). Thousand sperm were added into the media which including oocytes. Embryos were allowed to produce by IVF. The development of the embryos was followed until the 11th day, and the arrest, degeneration rates and alive embryos were established. The embryos reached 2, 4, 8, 16 cells stages and morula stage in the CM. While AMM had a negative effect on fertilization and embryo development, the most favourable effect was shown to be caused by CM in comparison with the other medias. These results have shown that the beneficial effects of CM in IVF would be a significant increase in the rate of fertility and development of embryos.

Mesenchymal stem cell conditioned media attenuates in vitro and ex vivo myocardial reperfusion injury

2011 ◽  
Vol 30 (1) ◽  
pp. 95-102 ◽  
Author(s):  
Denis Angoulvant ◽  
Fabrice Ivanes ◽  
René Ferrera ◽  
Phoebe G. Matthews ◽  
Serge Nataf ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Reperfusion Injury ◽  
Ex Vivo ◽  
Myocardial Reperfusion ◽  
Myocardial Reperfusion Injury ◽  
Conditioned Media
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