In Silico Identification of Transcription Factors Constituting the Regulatory Network that Drives Adipogenic Differentiation in Human Adipose-derived Stromal Cells In Vitro

Bone marrow-derived multipotent stromal cells (BMMSCs) represent an attractive therapeutic modality for cell therapy in type 2 diabetes mellitus (T2DM)-associated complications. T2DM changes the bone marrow environment; however, its effects on BMMSC properties remain unclear. The present study aimed at investigating select functions and differentiation of BMMSCs harvested from the T2DM microenvironment as potential candidates for regenerative medicine. BMMSCs were obtained from Zucker diabetic fatty (ZDF; an obese-T2DM model) rats and their lean littermates (ZL; controls), and cultured under normoglycemic conditions. The BMMSCs derived from ZDF animals were fewer in number, with limited clonogenicity (by 2-fold), adhesion (by 2.9-fold), proliferation (by 50%), migration capability (by 25%), and increased apoptosis rate (by 2.5-fold) compared to their ZL counterparts. Compared to the cultured ZL-BMMSCs, the ZDF-BMMSCs exhibited (i) enhanced adipogenic differentiation (increased number of lipid droplets by 2-fold; upregulation of the Pparg, AdipoQ, and Fabp genes), possibly due to having been primed to undergo such differentiation in vivo prior to cell isolation, and (ii) different angiogenesis-related gene expression in vitro and decreased proangiogenic potential after transplantation in nude mice. These results provided evidence that the T2DM environment impairs BMMSC expansion and select functions pertinent to their efficacy when used in autologous cell therapies.

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

Scientific Reports ◽

10.1038/s41598-021-85122-8 ◽

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.

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In Silico Identification and in Vitro Validation of Potential Cholestatic Compounds through 3D Ligand-Based Pharmacophore Modeling of BSEP Inhibitors

Chemical Research in Toxicology ◽

10.1021/tx5000393 ◽

2014 ◽

Vol 27 (5) ◽

pp. 873-881 ◽

Cited By ~ 20

Author(s):

Tina Ritschel ◽

Susanne M. A. Hermans ◽

Marieke Schreurs ◽

Jeroen J. M. W. van den Heuvel ◽

Jan B. Koenderink ◽

...

Keyword(s):

In Silico ◽

Pharmacophore Modeling ◽

In Silico Identification

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EST-based in silico identification and in vitro test of antimicrobial peptides in Brassica napus

BMC Genomics ◽

10.1186/s12864-015-1849-x ◽

2015 ◽

Vol 16 (1) ◽

Cited By ~ 4

Author(s):

Tao Ke ◽

Huihui Cao ◽

Junyan Huang ◽

Fan Hu ◽

Jin Huang ◽

...

Keyword(s):

Brassica Napus ◽

Antimicrobial Peptides ◽

In Silico ◽

In Vitro Test ◽

Vitro Test ◽

In Silico Identification

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In silico identification of viper phospholipaseA2 inhibitors: validation by in vitro, in vivo studies

Journal of Molecular Modeling ◽

10.1007/s00894-011-0994-7 ◽

2011 ◽

Vol 17 (12) ◽

pp. 3063-3073 ◽

Cited By ~ 9

Author(s):

Amit Nargotra ◽

Sujata Sharma ◽

Mohd Iqbal Alam ◽

Zabeer Ahmed ◽

Asha Bhagat ◽

...

Keyword(s):

In Silico ◽

In Vivo Studies ◽

In Silico Identification

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In silico identification and in vitro validation of nogalamycin N ‐oxide (NSC116555) as a potent anticancer compound against non–small‐cell lung cancer cells

Journal of Cellular Biochemistry ◽

10.1002/jcb.27605 ◽

2018 ◽

Vol 120 (3) ◽

pp. 3353-3361 ◽

Cited By ~ 2

Author(s):

Phongphat Obounchoey ◽

Lueacha Tabtimmai ◽

Praphasri Suphakun ◽

Kannika Thongkhao ◽

Chatchakorn Eurtivong ◽

...

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Cancer Cells ◽

Cell Lung Cancer ◽

In Silico ◽

Small Cell ◽

Small Cell Lung ◽

In Silico Identification ◽

Anticancer Compound

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Molecular Cloning of MAP Kinase Genes and In silico Identification of their Downstream Transcription Factors Involved in Pathogenesis of Karnal bunt(Tilletia indica) of Wheat

Journal of Proteomics & Bioinformatics ◽

10.4172/jpb.1000185 ◽

2011 ◽

Vol 04 (08) ◽

Author(s):

Atul K Gupta Anshita Goel

Keyword(s):

Transcription Factors ◽

Map Kinase ◽

Molecular Cloning ◽

In Silico ◽

Karnal Bunt ◽

Tilletia Indica ◽

In Silico Identification

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Bioreactor Processed Stromal Cell Seeding and Cultivation on Decellularized Pericardium Patches for Cardiovascular Use

Applied Sciences ◽

10.3390/app10165473 ◽

2020 ◽

Vol 10 (16) ◽

pp. 5473

Author(s):

Roman Matějka ◽

Miroslav Koňařík ◽

Jana Štěpanovská ◽

Jan Lipenský ◽

Jaroslav Chlupáč ◽

...

Keyword(s):

Cell Differentiation ◽

Stromal Cells ◽

Subcutaneous Fat ◽

Adipose Derived Stromal Cells ◽

Pulsatile Pressure ◽

Dynamic Cultivation ◽

Decellularized Tissue ◽

Static Conditions

(1) Background: Decellularized xenogeneic tissues are promising matrices for developing tissue-engineered cardiovascular grafts. In vitro recellularization of these tissues with stromal cells can provide a better in vivo remodelling and a lower thrombogenicity of the graft. The process of recellularization can be accelerated using a cultivation bioreactor simulating physiological conditions and stimuli. (2) Methods: Porcine pericardium was decellularized using a custom-built decellularization system with an optimized protocol. Autologous porcine adipose-derived stromal cells (PrASCs), isolated from the subcutaneous fat tissue, were used for recellularizing the decellularized pericardium. A custom cultivation bioreactor allowing the fixing of the decellularized tissue into a special cultivation chamber was created. The bioreactor maintained micro-perfusion and pulsatile pressure stimulation in order to promote the ingrowth of PrASCs inside the tissue and their differentiation. (3) Results: The dynamic cultivation promoted the ingrowth of cells into the decellularized tissue. Under static conditions, the cells penetrated only to the depth of 50 µm, whereas under dynamic conditions, the tissue was colonized up to 250 µm. The dynamic cultivation also supported the cell differentiation towards smooth muscle cells (SMCs). In order to ensure homogeneous cell colonization of the decellularized matrices, the bioreactor was designed to allow seeding of the cells from both sides of the tissue prior to the stimulation. In this case, the decellularized tissue was recolonized with cells within 5 days of dynamic cultivation. (4) Conclusions: Our newly designed dynamic bioreactor markedly accelerated the colonization of decellularized pericardium with ASCs and cell differentiation towards the SMC phenotype.

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