scholarly journals Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker

10.3791/57153 ◽  
2018 ◽  
Author(s):  
Jennifer Eom ◽  
Vaughan Feisst ◽  
Louis Ranjard ◽  
Kerry Loomes ◽  
Tanvi Damani ◽  
...  
Keyword(s):  
Adipogenic Differentiation ◽  
Mesenchymal Cell ◽  
Specific Marker ◽  
Differentiation Potential ◽  
Lineage Specific Marker

scholarly journals Metabolic Glycoengineering in hMSC-TERT as a Model for Skeletal Precursors by Using Modified Azide/Alkyne Monosaccharides

2021 ◽  
Vol 22 (6) ◽  
pp. 2820
Author(s):  
Stephan Altmann ◽  
Jürgen Mut ◽  
Natalia Wolf ◽  
Jutta Meißner-Weigl ◽  
Maximilian Rudert ◽  
...  
Keyword(s):  
Fluorescent Dyes ◽  
Adipogenic Differentiation ◽  
Culture Conditions ◽  
Surface Proteins ◽  
Differentiation Potential ◽  
Or Gene ◽  
Mannose Derivatives ◽  
Metabolic Glycoengineering ◽  
Scientific Questions

Metabolic glycoengineering enables a directed modification of cell surfaces by introducing target molecules to surface proteins displaying new features. Biochemical pathways involving glycans differ in dependence on the cell type; therefore, this technique should be tailored for the best results. We characterized metabolic glycoengineering in telomerase-immortalized human mesenchymal stromal cells (hMSC-TERT) as a model for primary hMSC, to investigate its applicability in TERT-modified cell lines. The metabolic incorporation of N-azidoacetylmannosamine (Ac4ManNAz) and N-alkyneacetylmannosamine (Ac4ManNAl) into the glycocalyx as a first step in the glycoengineering process revealed no adverse effects on cell viability or gene expression, and the in vitro multipotency (osteogenic and adipogenic differentiation potential) was maintained under these adapted culture conditions. In the second step, glycoengineered cells were modified with fluorescent dyes using Cu-mediated click chemistry. In these analyses, the two mannose derivatives showed superior incorporation efficiencies compared to glucose and galactose isomers. In time-dependent experiments, the incorporation of Ac4ManNAz was detectable for up to six days while Ac4ManNAl-derived metabolites were absent after two days. Taken together, these findings demonstrate the successful metabolic glycoengineering of immortalized hMSC resulting in transient cell surface modifications, and thus present a useful model to address different scientific questions regarding glycosylation processes in skeletal precursors.

scholarly journals Bone marrow stromal cells from MDS and AML patients show increased adipogenic potential with reduced Delta-like-1 expression

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marie-Theresa Weickert ◽  
Judith S. Hecker ◽  
Michèle C. Buck ◽  
Christina Schreck ◽  
Jennifer Rivière ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Fate ◽  
Stromal Cells ◽  
Adipogenic Differentiation ◽  
Cell Types ◽  
Bone Marrow Niche ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Bm Niche

AbstractMyelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are clonal hematopoietic stem cell disorders with a poor prognosis, especially for elderly patients. Increasing evidence suggests that alterations in the non-hematopoietic microenvironment (bone marrow niche) can contribute to or initiate malignant transformation and promote disease progression. One of the key components of the bone marrow (BM) niche are BM stromal cells (BMSC) that give rise to osteoblasts and adipocytes. It has been shown that the balance between these two cell types plays an important role in the regulation of hematopoiesis. However, data on the number of BMSC and the regulation of their differentiation balance in the context of hematopoietic malignancies is scarce. We established a stringent flow cytometric protocol for the prospective isolation of a CD73+ CD105+ CD271+ BMSC subpopulation from uncultivated cryopreserved BM of MDS and AML patients as well as age-matched healthy donors. BMSC from MDS and AML patients showed a strongly reduced frequency of CFU-F (colony forming unit-fibroblast). Moreover, we found an altered phenotype and reduced replating efficiency upon passaging of BMSC from MDS and AML samples. Expression analysis of genes involved in adipo- and osteogenic differentiation as well as Wnt- and Notch-signalling pathways showed significantly reduced levels of DLK1, an early adipogenic cell fate inhibitor in MDS and AML BMSC. Matching this observation, functional analysis showed significantly increased in vitro adipogenic differentiation potential in BMSC from MDS and AML patients. Overall, our data show BMSC with a reduced CFU-F capacity, and an altered molecular and functional profile from MDS and AML patients in culture, indicating an increased adipogenic lineage potential that is likely to provide a disease-promoting microenvironment.

scholarly journals Similar Features, Different Behaviors: A Comparative In Vitro Study of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament

2021 ◽  
Vol 11 (8) ◽  
pp. 738
Author(s):  
Melissa D. Mercado-Rubio ◽  
Erick Pérez-Argueta ◽  
Alejandro Zepeda-Pedreguera ◽  
Fernando J. Aguilar-Ayala ◽  
Ricardo Peñaloza-Cuevas ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Periodontal Ligament ◽  
Adipogenic Differentiation ◽  
In Vitro Study ◽  
Mrna Levels ◽  
Dental Tissue ◽  
Periodontal Ligament Stem Cells ◽  
Differentiation Potential

Dental tissue-derived mesenchymal stem cells (DT-MSCs) are a promising resource for tissue regeneration due to their multilineage potential. Despite accumulating data regarding the biology and differentiation potential of DT-MSCs, few studies have investigated their adipogenic capacity. In this study, we have investigated the mesenchymal features of dental pulp stem cells (DPSCs), as well as the in vitro effects of different adipogenic media on these cells, and compared them to those of periodontal ligament stem cells (PLSCs) and dental follicle stem cells (DFSCs). DFSC, PLSCs, and DPSCs exhibit similar morphology and proliferation capacity, but they differ in their self-renewal ability and expression of stemness markers (e.g OCT4 and c-MYC). Interestingly, DFSCs and PLSCs exhibited more lipid accumulation than DPSCs when induced to adipogenic differentiation. In addition, the mRNA levels of adipogenic markers (PPAR, LPL, and ADIPOQ) were significantly higher in DFSCs and PLSCs than in DPSCs, which could be related to the differences in the adipogenic commitment in those cells. These findings reveal that the adipogenic capacity differ among DT-MSCs, features that might be advantageous to increasing our understanding about the developmental origins and regulation of adipogenic commitment.

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.

220 CpG METHYLATION PROFILE OF ADIPOGENIC AND ENDOTHELIAL GENE PROMOTERS IN HUMAN ADIPOSE STEM CELLS SUGGESTS A RESTRICTIVE ENDOTHELIAL DIFFERENTIATION POTENTIAL

2007 ◽  
Vol 19 (1) ◽  
pp. 227
Author(s):  
A. C. Boquest ◽  
A. Noer ◽  
A. L. Sørensen ◽  
K. Vekterud ◽  
P. Collas
Keyword(s):  
Stem Cells ◽  
Adipogenic Differentiation ◽  
Cpg Methylation ◽  
Methylation Profile ◽  
Adipose Stem Cells ◽  
Endothelial Differentiation ◽  
Sequencing Analysis ◽  
Differentiation Potential ◽  
Undifferentiated Cells ◽  
Endothelial Lineage

Mesenchymal stem cells (MSCs) have received intense research interest due to their perceived potential application in regenerative medicine; nevertheless, MSCs are primarily restricted to form mesodermal cell types. Adipose stem cells (ASCs) with a CD34+ CD105+ CD45– CD31– immunophenotype can be obtained in an uncultured state with high purity from the stromal vascular fraction of human liposuction material (Boquest et al. 2005 Mol. Biol. Cell 16, 1131–1141). While ASCs differentiate readily into adipocytes, their endothelial lineage commitment has been scarcely reported, and controversy remains regarding ASC contribution to vascularization. To address the epigenetic commitment of ASCs to adipogenic and endothelial lineages, we carried out a bisulfite sequencing analysis of CpG methylation in the promoters of adipogenic (LEP, PPARG2, FABP4, LPL), endothelial (CD31, CD144), and myogenic (MYOG) genes in freshly isolated and in clonal ASC cultures in relation to gene expression and differentiation potential. Uncultured ASCs display mosaic hypomethylation of adipogenic promoters, in contrast to MYOG, CD31, or CD144 which are methylated (Noer et al. 2006 Mol. Biol. Cell 17, in press). Nevertheless, CpG methylation does not reflect transcriptional status of these genes in undifferentiated cells. Culture and adipogenic differentiation of ASCs maintains the hypomethylated profile of adipogenic promoters and the hypermethylation of non-adipogenic promoters. Endothelial stimulation of ASCs in methylcellulose elicits tubule-like networks, up-regulation of CD31 and CD144, and restrictive induction of a CD31+ CD144+ immunophenotype. Discrete and lineage-specific changes in CpG methylation in the CD31 and CD144 promoters take place but no global demethylation that marks endothelial cells occurs. Promoters not involved in endothelial differentiation retain a methylation profile characteristic of undifferentiated cells. Hypermethylation of CD31 and CD144 suggests a restricted commitment of ASCs to the endothelial lineage. This contrasts with hypomethylation of adipogenic promoters which reflects a propensity toward adipogenic differentiation. Despite the up-regulation of lineage-specific transcripts, overall maintenance of promoter methylation after adipogenic, osteogenic, and endothelial differentiation suggests the maintenance of an epigenetic signature characteristic of undifferentiated cells. Analysis of CpG methylation at lineage-specific promoters should provide a robust assessment of epigenetic commitment of stem cells to a specific lineage.

scholarly journals The Effect of Early Rounds of ex vivo Expansion and Cryopreservation on the Adipogenic Differentiation Capacity of Adipose-Derived Stromal/Stem Cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
C. Durandt ◽  
C. Dessels ◽  
C. da Silva ◽  
C. Murdoch ◽  
M. S. Pepper
Keyword(s):  
Stem Cells ◽  
Ex Vivo ◽  
Adipogenic Differentiation ◽  
Isolated Cells ◽  
Differentiation Potential ◽  
Differentiation Capacity ◽  
Surface Marker Expression ◽  
Cd36 Expression ◽  
The Impact ◽  
Stromal Stem Cells

Abstract Multipotent adipose-derived stromal/stem cells (ASCs) are candidates for use in cellular therapies for the treatment of a variety of conditions/diseases. Ex vivo expansion of freshly isolated ASCs may be necessary prior to clinical application to ensure that clinically relevant cell numbers are administered during treatment. In addition, cryopreserving cells at early passages allows for storage of freshly isolated cells for extended periods of time before expanding these cells for clinical usage. There are however several concerns that these laboratory-based procedures may alter the characteristics of the cells and in so doing decrease their regenerative potential. In this study we report on the impact of early rounds of cryopreservation (P0) and ex vivo expansion (P0 to P5) on the phenotypic characteristics and adipogenic differentiation potential of ASCs. Our results show that ASCs that upregulate CD36 expression during adipogenic differentiation gradually decrease with increasing expansion rounds. The consequent decrease in adipogenic differentiation capacity was evident in both gene expression and flow cytometry-based phenotypic studies. Successive rounds of expansion did not however alter cell surface marker expression of the cells. We also show that early cryopreservation of ASCs (at P0) does not affect the adipogenic differentiation potential of the cells.

scholarly journals Mesenchymal Progenitors Derived from Different Locations in Long Bones Display Diverse Characteristics

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Weiguang Lu ◽  
Bo Gao ◽  
Jing Fan ◽  
Pengzhen Cheng ◽  
Yaqian Hu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Adipogenic Differentiation ◽  
Osteoclast Formation ◽  
Long Bones ◽  
Differentiation Potential ◽  
Mesenchymal Progenitors ◽  
Multiple Differentiation ◽  
Osteoclastic Differentiation ◽  
Proliferation Assays

Mesenchymal progenitors within bone marrow have multiple differentiation potential and play an essential role in the maintenance of adult skeleton homeostasis. Mesenchymal progenitors located in bone regions other than the bone marrow also display bone-forming properties. However, owing to the differences in each distinct microenvironment, the mesenchymal characteristics of skeletal progenitor cells within different regions of long bones may show some differences. In order to clearly elucidate these differences, we performed a comparative study on mesenchymal progenitors from different regions of long bones. Here, we isolated mesenchymal progenitors from the periosteum, endosteum, and bone marrow of rat long bones. The three groups exhibited similar cellular morphologies and expressed the typical surface markers associated with mesenchymal stem cells. Interestingly, after cell proliferation assays and bidirectional differentiation analysis, periosteal mesenchymal progenitors showed a higher proliferative ability and adipogenic differentiation potential. In contrast, endosteal mesenchymal progenitors were more prone to osteogenic differentiation. Using in vitro osteoclast culture systems, conditioned media from different mesenchymal progenitor cultures were used to induce osteoclastic differentiation. Osteoclast formation was found to be significantly promoted by the secretion of RANKL and IL-6 by endosteal progenitors. Overall, our results provide strong evidence for the importance of selecting the appropriate source of skeletal progenitors for applications in future skeleton regeneration therapies.

scholarly journals “The preadipocyte factor” DLK1 marks adult mouse adipose tissue residing vascular cells that lack in vitro adipogenic differentiation potential

FEBS Letters ◽  
2009 ◽  
Vol 583 (17) ◽  
pp. 2947-2953 ◽  
Author(s):  
Ditte Caroline Andersen ◽  
Line Jensen ◽  
Henrik Daa Schrøder ◽  
Charlotte Harken Jensen
Keyword(s):  
Adipose Tissue ◽  
Adult Mouse ◽  
Adipogenic Differentiation ◽  
Vascular Cells ◽  
Differentiation Potential
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