scholarly journals Gene Expression of Candidate Genes Involved in Fat Metabolism During In vitro Adipogenic Differentiation of Bovine Mesenchymal Stem Cell

2010 ◽  
Vol 52 (4) ◽  
pp. 265-270 ◽  
Author(s):  
Sung-Kon Kim ◽  
Nam-Kuk Kim ◽  
Du-Hak Yoon ◽  
Tae-Hun Kim ◽  
Boo-Keun Yang ◽  
...  
2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Francesco Da Ros ◽  
Luca Persano ◽  
Dario Bizzotto ◽  
Mariagrazia Michieli ◽  
Paola Braghetta ◽  
...  

Abstract Background Dissection of mechanisms involved in the regulation of bone marrow microenvironment through cell–cell and cell–matrix contacts is essential for the detailed understanding of processes underlying bone marrow activities both under physiological conditions and in hematologic malignancies. Here we describe Emilin-2 as an abundant extracellular matrix component of bone marrow stroma. Methods Immunodetection of Emilin-2 was performed in bone marrow sections of mice from 30 days to 6 months of age. Emilin-2 expression was monitored in vitro in primary and mesenchymal stem cell lines under undifferentiated and adipogenic conditions. Hematopoietic stem cells and progenitors in bone marrow of 3- to 10-month-old wild-type and Emilin-2 null mice were analyzed by flow cytometry. Results Emilin-2 is deposited in bone marrow extracellular matrix in an age-dependent manner, forming a meshwork that extends from compact bone boundaries to the central trabecular regions. Emilin-2 is expressed and secreted by both primary and immortalized bone marrow mesenchymal stem cells, exerting an inhibitory action in adipogenic differentiation. In vivo Emilin-2 deficiency impairs the frequency of hematopoietic stem/progenitor cells in bone marrow during aging. Conclusion Our data provide new insights in the contribution of bone marrow extracellular matrix microenvironment in the regulation of stem cell niches and hematopoietic progenitor differentiation.


2019 ◽  
Vol 7 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Vivi Sofia ◽  
Ellyza Nasrul ◽  
Menkher Manjas ◽  
Gusti Revilla

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).


Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 268
Author(s):  
Kihae Ra ◽  
Hyun Ju Oh ◽  
Eun Young Kim ◽  
Sung Keun Kang ◽  
Jeong Chan Ra ◽  
...  

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.


Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 341
Author(s):  
Tobias Russell ◽  
Abdulla Watad ◽  
Charlie Bridgewood ◽  
Hannah Rowe ◽  
Almas Khan ◽  
...  

Objective: The spondylarthritides (SpA) are intimately linked to new bone formation and IL-17A and TNF pathways. We investigated spinal soft tissue and bone mesenchymal stem cell (MSC) responses to IL-17A and TNF, including their osteogenesis, adipogenesis, and stromal supportive function and ability to support lymphocyte recruitment. Methods: Normal spinal peri-entheseal bone (PEB) and entheseal soft tissue (EST) were characterized for MSCs by immunophenotypic, osteogenic, chondrogenic, and adipogenic differentiation criteria. Functional and gene transcriptomic analysis was carried out on undifferentiated, adipo- differentiated, and osteo-differentiated MSCs. The enthesis C-C Motif Chemokine Ligand 20-C-C Motif Chemokine Receptor 6 (CCL20-CCR6) axis was investigated at transcript and protein levels to ascertain whether entheseal MSCs influence local immune cell populations. Results: Cultured MSCs from both PEB and EST displayed a tri-lineage differentiation ability. EST MSCs exhibited 4.9-fold greater adipogenesis (p < 0.001) and a 3-fold lower osteogenic capacity (p < 0.05). IL-17A induced greater osteogenesis in PEB MSCs compared to EST MSCs. IL-17A suppressed adipogenic differentiation, with a significant decrease in fatty acid-binding protein 4 (FABP4), peroxisome proliferator-activated receptor gamma (PPARγ), Cell Death Inducing DFFA Like Effector C (CIDEC), and Perilipin-1 (PLIN1). IL-17A significantly increased the CCL20 transcript (p < 0.01) and protein expression (p < 0.001) in MSCs supporting a role in type 17 lymphocyte recruitment. Conclusions: Normal spinal enthesis harbors resident MSCs with different in vitro functionalities in bone and soft tissue, especially in response to IL-17A, which enhanced osteogenesis and CCL20 production and reduced adipogenesis compared to unstimulated MSCs. This MSC-stromal-enthesis immune system may be a hitherto unappreciated mechanism of “fine tuning” tissue repair responses at the enthesis in health and could be relevant for SpA understanding.


2019 ◽  
Vol 7 (4) ◽  
pp. 543-548 ◽  
Author(s):  
Vivi Sofia ◽  
Ellyza Nasrul ◽  
Menkher Manjas ◽  
Gusti Revilla

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).


Author(s):  
Sara Svensson ◽  
Michael Palmer ◽  
Johan Svensson ◽  
Anna Johansson ◽  
Håkan Engqvist ◽  
...  

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.


2021 ◽  
Vol 49 (2) ◽  
pp. 332-339
Author(s):  
Hideyuki Shirasawa ◽  
Noboru Matsumura ◽  
Masaki Yoda ◽  
Kazumasa Okubo ◽  
Masayuki Shimoda ◽  
...  

Background: The infiltration of fat tissue into skeletal muscle, a condition referred to as muscle fatty infiltration or fatty degeneration, is regarded as an irreversible event that significantly compromises the motor function of skeletal muscle. Purpose: To investigate the effect of retinoic acid receptor (RAR) agonists in suppressing the adipogenic differentiation of fibroadipogenic progenitors (FAPs) in vitro and fatty infiltration after rotator cuff tear in mice. Study Design: Controlled laboratory study. Methods: FAPs isolated from mouse skeletal muscle were cultured in adipogenic differentiation medium in the presence or absence of an RAR agonist. At the end of cell culture, adipogenic differentiation was evaluated by gene expression analysis and oil red O staining. A mouse model of fatty infiltration—which includes the resection of the rotator cuff, removal of the humeral head, and denervation the supraspinatus muscle—was used to induce fatty infiltration in the supraspinatus muscle. The mice were orally or intramuscularly administered with an RAR agonist after the surgery. Muscle fatty infiltration was evaluated by histology and gene expression analysis. Results: RAR agonists effectively inhibited the adipogenic differentiation of FAPs in vitro. Oral and intramuscular administration of RAR agonists suppressed the development of muscle fatty infiltration in the mice after rotator cuff tear. In accordance, we found a significant decrease in the number of intramuscular fat cells and suppressed expression in adipogenic markers. RAR agonists also increased the expression of the transcripts for collagens; however, an accumulation of collagenous tissues was not histologically evident in the present model. Conclusion: Muscle fatty infiltration can be alleviated by RAR agonists through suppressing the adipogenic differentiation of FAPs. The results also suggest that RAR agonists are potential therapeutic agents for treating patients who are at risk of developing muscle fatty infiltration. The consequence of the increased expression of collagen transcripts by RAR agonists needs to be clarified. Clinical Relevance: RAR agonists can be used to prevent the development of muscle fatty infiltration after rotator cuff tear. Nevertheless, further studies are mandatory in a large animal model to examine the safety and efficacy of intramuscular injection of RAR agonists.


Sign in / Sign up

Export Citation Format

Share Document