scholarly journals Effect of 20(S)-Hydroxycholesterol on Multilineage Differentiation of Mesenchymal Stem Cells Isolated from Compact Bones in Chicken

Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1360
Author(s):  
Roshan Adhikari ◽  
Chongxiao Chen ◽  
Woo Kyun Kim
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Health ◽  
Hedgehog Signaling ◽  
Myogenic Differentiation ◽  
Adipogenic Differentiation ◽  
Bioactive Molecules ◽  
Marker Genes ◽  
Human Mscs ◽  
Differentiation Potential

Bone health and body weight gain have significant economic and welfare importance in the poultry industry. Mesenchymal stem cells (MSCs) are common progenitors of different cell lineages such as osteoblasts, adipocytes, and myocytes. Specific oxysterols have shown to be pro-osteogenic and anti-adipogenic in mouse and human MSCs. To determine the effect of 20(S)-hydroxycholesterol (20S) on osteogenic, adipogenic, and myogenic differentiation in chicken, mesenchymal stem cells isolated from compact bones of broiler chickens (cBMSCs) were subjected to various doses of 20S, and markers of lineage-specific mRNA were analyzed using real-time PCR and cell cytochemistry. Further studies were conducted to evaluate the molecular mechanisms involved in lineage-specific differentiation pathways. Like human and mouse MSCs, 20S oxysterol expressed pro-osteogenic, pro-myogenic, and anti-adipogenic differentiation potential in cBMSCs. Moreover, 20(S)-Hydroxycholesterol induced markers of osteogenic genes and myogenic regulatory factors when exposed to cBMSCs treated with their specific medium. In contrast, 20S oxysterol suppressed expression of adipogenic marker genes when exposed to cBMSCs treated with OA, an adipogenic precursor of cBMSCs. To elucidate the molecular mechanism by which 20S exerts its differentiation potential in all three lineages, we focused on the hedgehog signaling pathway. The hedgehog inhibitor, cyclopamine, completely reversed the effect of 20S induced expression of osteogenic and anti-adipogenic mRNA. However, there was no change in the mRNA expression of myogenic genes. The results showed that 20S oxysterol promotes osteogenic and myogenic differentiation and decreases adipocyte differentiation of cBMSCs. This study also showed that the induction of osteogenesis and adipogenesis inhibition in cBMSCs by 20S is mediated through the hedgehog signaling mechanism. The results indicated that 20(S) could play an important role in the differentiation of chicken-derived MSCs and provided the theory basis on developing an intervention strategy to regulate skeletal, myogenic, and adipogenic differentiation in chicken, which will contribute to improving chicken bone health and meat quality. The current results provide the rationale for the further study of regulatory mechanisms of bioactive molecules on the differentiation of MSCs in chicken, which can help to address skeletal health problems in poultry.

scholarly journals Adipogenic differentiation potential of porcine bone marrow mesenchymal stem cells grown in different basal media formulations

2020 ◽  
Vol 11 (4) ◽  
pp. 7108-7120
Author(s):  
Sangeetha Kannan ◽  
Jyotirmoy Ghosh ◽  
Sujoy K. Dhara
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Significant Role ◽  
Culture Media ◽  
Adipogenic Differentiation ◽  
Marker Genes ◽  
Differentiation Potential ◽  
Basal Media ◽  
Adipogenic Marker

Bone marrow-derived porcine mesenchymal stem cells (pMSC) are precursors of multiple lineage cells. However, the differentiation potential of pMSC might vary due to the culture media in which they are isolated and grown. In this study the effects of αMEM, aDMEM, M199, αMEM/M199, aDMEM/M199 and αMEM/aDMEM was determined on pMSC expression of adipogenic marker genes PPARγ, C/EBPα and ApN at passage 5 and 10; and differentiation potential of pMSC at passage 5. Relative expressions of the genes were determined by qPCR assay. Adipogenic differentiation progress was determined by microscopic evaluation and accumulation of lipid vesicles in cells after day 21 differentiation was confirmed by Oil Red-O staining. Results indicated varying expressions of adipogenic marker genes at passage 5 and 10 in pMSC grown in different culture media. Adipogenic differentiation characteristics also varied in different culture media, grown cells depending on the expression of adipogenic marker genes. Classification in order of differentiation potential revealed that the pMSC grown in M199 and αMEM/M199 was the best, followed by aDMEM/M199 and αMEM/aDMEM the intermediate and αMEM and aDMEM the least suitable media. Effect of media interaction analysis indicated that M199 media might have played a significant role in up-keeping of the desired marker gene expression and adipogenic differentiation potential of the pMSC. In conclusion, adipogenic differentiation potential of pMSC varied due to the differences in basal media composition and the M199 played a significant role in skewing pMSC towards adipogenic lineage.

scholarly journals Epigenetic Regulation of Mesenchymal Stem Cells: A Focus on Osteogenic and Adipogenic Differentiation

2011 ◽  
Vol 2011 ◽  
pp. 1-18 ◽  
Author(s):  
Chad M. Teven ◽  
Xing Liu ◽  
Ning Hu ◽  
Ni Tang ◽  
Stephanie H. Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Epigenetic Regulation ◽  
Adult Stem Cells ◽  
Es Cells ◽  
Adipogenic Differentiation ◽  
Embryonic Stem ◽  
Cell Types ◽  
Differentiation Potential ◽  
Msc Differentiation

Stem cells are characterized by their capability to self-renew and terminally differentiate into multiple cell types. Somatic or adult stem cells have a finite self-renewal capacity and are lineage-restricted. The use of adult stem cells for therapeutic purposes has been a topic of recent interest given the ethical considerations associated with embryonic stem (ES) cells. Mesenchymal stem cells (MSCs) are adult stem cells that can differentiate into osteogenic, adipogenic, chondrogenic, or myogenic lineages. Owing to their ease of isolation and unique characteristics, MSCs have been widely regarded as potential candidates for tissue engineering and repair. While various signaling molecules important to MSC differentiation have been identified, our complete understanding of this process is lacking. Recent investigations focused on the role of epigenetic regulation in lineage-specific differentiation of MSCs have shown that unique patterns of DNA methylation and histone modifications play an important role in the induction of MSC differentiation toward specific lineages. Nevertheless, MSC epigenetic profiles reflect a more restricted differentiation potential as compared to ES cells. Here we review the effect of epigenetic modifications on MSC multipotency and differentiation, with a focus on osteogenic and adipogenic differentiation. We also highlight clinical applications of MSC epigenetics and nuclear reprogramming.

Effect of Substrate Elasticity on In Vitro Aging of Human Mesenchymal Stem Cells

2012 ◽  
Vol 1498 ◽  
pp. 39-45
Author(s):  
Courtney E. LeBlon ◽  
Caitlin R. Fodor ◽  
Tony Zhang ◽  
Xiaohui Zhang ◽  
Sabrina S. Jedlicka
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Elastic Modulus ◽  
Myogenic Differentiation ◽  
Human Mesenchymal Stem Cells ◽  
Immunofluorescent Staining ◽  
Differentiation Potential ◽  
Gene And Protein Expression ◽  
The Impact

ABSTRACTHuman mesenchymal stem cells (hMSCs) were routinely cultured on tissue-culture polystyrene (TCPS) to investigate the in vitro aging and cell stiffening. hMSCs were also cultured on thermoplastic polyurethane (TPU), which is a biocompatible polymer with an elastic modulus of approximately 12.9MPa, to investigate the impact of substrate elastic modulus on cell stiffening and differentiation potential. Cells were passaged over several generations on each material. At each passage, cells were subjected to osteogenic and myogenic differentiation. Local cell elastic modulus was measured at every passage using atomic force microscopy (AFM) indentation. Gene and protein expression was examined using qRT-PCR and immunofluorescent staining, respectively, for osteogenic and myogenic markers. Results show that the success of myogenic differentiation is highly reliant on the elastic modulus of the undifferentiated cells. The success of osteogenic differentiations is most likely somewhat dependent on the cell elastic modulus, as differentiations were more successful in earlier passages, when cells were softer.

scholarly journals Myogenic differentiation potential of chicken mesenchymal stem cells from bone marrow

2021 ◽  
Author(s):  
Zhen Zhou ◽  
Changbin Zhao ◽  
Bolin Cai ◽  
Manting Ma ◽  
Shaofen Kong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Muscle Development ◽  
Myogenic Differentiation ◽  
Horse Serum ◽  
Growth Characteristic ◽  
Normal Growth ◽  
Receptor Interaction ◽  
Differentiation Potential ◽  
Differential Adhesion

Abstract Background: Mesenchymal stem cells (MSCs) have the potential to multilineage differentiation, which can be used for a good model to provide critical insight of chicken muscle development. Differential adhesion method is one of the commonest methods to isolate MSCs based on the ability of plastic adhesion. 5-azacytidine (5-Aza), dexamethasone (DXMS), hydrocortisone (HC) and horse serum had been proved the potential to induce the myogenic differentiation of MSCs. However, the myogenic differentiation of MSCs is still poorly understood in chicken. In present study, we isolated chicken mesenchymal stem cells (cMSCs) from bone using 4-hour differential adhesion method and analyzed the myogenic effect of cMSCs treated with different method based on 5-Aza, DXMS, HC and horse serum.Results: cMSCs isolated by 4-hour differential adhesion method expressed MSCs special surface markers and presented normal growth characteristic. cMSCs showed great potential of myogenic differentiation by the treatment of 5-Aza and horse serum. RNA-sequence, GO and KGEE enrichment analysis revealed that this effect might be based on demethylation of 5-Aza and ECM-receptor interaction, focal adhesion, PI3K-Akt, p53, TGF-beta signaling pathways. Moreover, DXMS, HC and horse serum also presented potential of myogenic differentiation, but the effect was not as good as 5-Aza and horse serum method.Conclusions: cMSCs showed potential of myogenic differentiation by the treatment of 5-Aza and horse serum or DXMS, HC and horse serum.

scholarly journals Effects of Severe Hypoxia on Bone Marrow Mesenchymal Stem Cells Differentiation Potential

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Claudia Cicione ◽  
Emma Muiños-López ◽  
Tamara Hermida-Gómez ◽  
Isaac Fuentes-Boquete ◽  
Silvia Díaz-Prado ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Molecular Mechanisms ◽  
Control Point ◽  
Severe Hypoxia ◽  
Marker Genes ◽  
Differentiation Potential ◽  
Differentiation Capacity ◽  
Hypoxic Conditions

Background. The interests in mesenchymal stem cells (MSCs) and their application in cell therapy have resulted in a better understanding of the basic biology of these cells. Recently hypoxia has been indicated as crucial for complete chondrogenesis. We aimed at analyzing bone marrow MSCs (BM-MSCs) differentiation capacity under normoxic and severe hypoxic culture conditions.Methods. MSCs were characterized by flow cytometry and differentiated towards adipocytes, osteoblasts, and chondrocytes under normoxic or severe hypoxic conditions. The differentiations were confirmed comparing each treated point with a control point made of cells grown in DMEM and fetal bovine serum (FBS).Results. BM-MSCs from the donors displayed only few phenotypical differences in surface antigens expressions. Analyzing marker genes expression levels of the treated cells compared to their control point for each lineage showed a good differentiation in normoxic conditions and the absence of this differentiation capacity in severe hypoxic cultures.Conclusions. In our experimental conditions, severe hypoxia affects thein vitrodifferentiation potential of BM-MSCs. Adipogenic, osteogenic, and chondrogenic differentiations are absent in severe hypoxic conditions. Our work underlines that severe hypoxia slows cell differentiation by means of molecular mechanisms since a decrease in the expression of adipocyte-, osteoblast-, and chondrocyte-specific genes was observed.

scholarly journals Mechanisms Underlying the Osteo- and Adipo-Differentiation of Human Mesenchymal Stem Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Yu Zhang ◽  
Dilaware Khan ◽  
Julia Delling ◽  
Edda Tobiasch
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipogenic Differentiation ◽  
Human Mesenchymal Stem Cells ◽  
Human Mscs ◽  
Industrialized Countries ◽  
Tight Control ◽  
Age Related ◽  
Donor Cells ◽  
Immunosuppressive Microenvironment

Human mesenchymal stem cells (hMSCs) are considered a promising cell source for regenerative medicine, because they have the potential to differentiate into a variety of lineages among which the mesoderm-derived lineages such adipo- or osteogenesis are investigated best. Human MSCs can be harvested in reasonable to large amounts from several parts of the patient’s body and due to this possible autologous origin, allorecognition can be avoided. In addition, even in allogenic origin-derived donor cells, hMSCs generate a local immunosuppressive microenvironment, causing only a weak immune reaction. There is an increasing need for bone replacement in patients from all ages, due to a variety of reasons such as a new recreational behavior in young adults or age-related diseases. Adipogenic differentiation is another interesting lineage, because fat tissue is considered to be a major factor triggering atherosclerosis that ultimately leads to cardiovascular diseases, the main cause of death in industrialized countries. However, understanding the differentiation process in detail is obligatory to achieve a tight control of the process for future clinical applications to avoid undesired side effects. In this review, the current findings for adipo- and osteo-differentiation are summarized together with a brief statement on first clinical trials.

scholarly journals The Influence of Aging on the Regenerative Potential of Human Adipose Derived Mesenchymal Stem Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Monika Marędziak ◽  
Krzysztof Marycz ◽  
Krzysztof A. Tomaszewski ◽  
Katarzyna Kornicka ◽  
Brandon Michael Henry
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipogenic Differentiation ◽  
Population Doubling ◽  
Osteogenic Potential ◽  
Population Doubling Time ◽  
Joint Diseases ◽  
Differentiation Potential ◽  
Age Related

Tissue regeneration using human adipose derived mesenchymal stem cells (hASCs) has significant potential as a novel treatment for many degenerative bone and joint diseases. Previous studies have established that age negatively affects the proliferation status and the osteogenic and chondrogenic differentiation potential of mesenchymal stem cells. The aim of this study was to assess the age-related maintenance of physiological function and differentiation potential of hASCs in vitro. hASCs were isolated from patients of four different age groups: (1) >20 years (n=7), (2) >50 years (n=7), (3) >60 years (n=7), and (4) >70 years (n=7). The hASCs were characterized according to the number of fibroblasts colony forming unit (CFU-F), proliferation rate, population doubling time (PDT), and quantified parameters of adipogenic, chondrogenic, and osteogenic differentiation. Compared to younger cells, aged hASCs had decreased proliferation rates, decreased chondrogenic and osteogenic potential, and increased senescent features. A shift in favor of adipogenic differentiation with increased age was also observed. As many bone and joint diseases increase in prevalence with age, it is important to consider the negative influence of age on hASCs viability, proliferation status, and multilineage differentiation potential when considering the potential therapeutic applications of hASCs.

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