Wnt Signaling Regulates the Lymphatic Endothelial Transdifferentiation of Adipose-Derived Stromal Cells In Vitro

2021 ◽  
Author(s):  
Nian Zhang ◽  
Liru Hu ◽  
Jiyuan Liu ◽  
Wenbin Yang ◽  
Ye Li ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Stromal Cells ◽  
Adipose Derived Stromal Cells

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

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.

Exosome derived from murine adipose-derived stromal cells: Neuroprotective effect on in vitro model of amyotrophic lateral sclerosis

2016 ◽  
Vol 340 (1) ◽  
pp. 150-158 ◽  
Author(s):  
Roberta Bonafede ◽  
Ilaria Scambi ◽  
Daniele Peroni ◽  
Valentina Potrich ◽  
Federico Boschi ◽  
...  
Keyword(s):  
Amyotrophic Lateral Sclerosis ◽  
Stromal Cells ◽  
In Vitro Model ◽  
Neuroprotective Effect ◽  
Adipose Derived Stromal Cells ◽  
Vitro Model ◽  
Lateral Sclerosis

Chondrogenesis by Co-culture of Adipose-Derived Stromal Cells and Chondrocytes In Vitro

2012 ◽  
Vol 53 (6) ◽  
pp. 492-497 ◽  
Author(s):  
Xiaojie Lv ◽  
Guangdong Zhou ◽  
Xia Liu ◽  
Huxian Liu ◽  
Junnan Chen ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Adipose Derived Stromal Cells

Implications of adipose-derived stromal cells in a 3D culture system for osteogenic differentiation: an in vitro and in vivo investigation

2013 ◽  
Vol 13 (1) ◽  
pp. 32-43 ◽  
Author(s):  
Francis H. Shen ◽  
Brian C. Werner ◽  
Haixiang Liang ◽  
Hulan Shang ◽  
Ning Yang ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Culture System ◽  
3D Culture ◽  
Adipose Derived Stromal Cells ◽  
3D Culture System

scholarly journals Towards a Standardization and Characterization of Clinical Grade Adipose-Derived Stromal Cells

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5013-5013
Author(s):  
Chiara Cugno ◽  
Rita Calzone ◽  
Giusy Gentilcore ◽  
Heba Sidahmed ◽  
Asma Al-Sulaiti ◽  
...  
Keyword(s):  
Mirna Expression ◽  
Stromal Cells ◽  
Conflicts Of Interest ◽  
Cell Yield ◽  
Population Doubling ◽  
Color Flow ◽  
Single Donor ◽  
Adipose Derived Stromal Cells

INTRODUCTION Mesenchymal Stromal Cells (MSCs) are multipotent cells with regenerative, anti-inflammatory, immunomodulatory and anti-tumorigenic properties, which are readily isolated from aspirates of adipose tissue (AT) due to their high availability. Compared with alternative sources, AT provides high cell yield. Despite the numerous clinical trials in autologous and allogeneic settings for various diseases worldwide, there are no US FDA-approved MSC-based products. Challenges for clinical translation of MSC-based therapies largely lie in the sourcing, production and basic characterization of such products (Camilleri et al, 2016; Mendicino et al, 2014). Given the known donor-related variability (sex, age, liposuction sites, BMI, etc.), there is still ample room to: better standardize in vitro manufacturing processesrefine the characterization of clinical products. METHODS Seven samples of lipoaspirate were processed. AT manual digestion was compared with an automated collagenase-based enzymatic digestion by the Cytori Celution instrument, a closed system regulated as a medical device by EC for the production of adipose-derived stem cells-enriched products. Total cell yield, cell harvest at P0, P1 and P2, Population Doubling Level (PDL) and Doubling Time (DT) were evaluated. The obtained Adipose-derived Stromal Cells (ADSCs) were characterized for: Expression of 361 surface markers (BioLegend LegendScreen Human PE Kit), including 117 additional antigens compared to other reported screens. Cytori-isolated ADSCs from three donors for 6 total conditions (3 P2, 2 P3 and 1 IFN-g-primed P2) were screened. A broader characterization of fresh isolates (P0) as well as (P3) from the same donors was performed with a 20-color flow cytometric panel. Data were analyzed with FlowJo software.miRNA expression was evaluated using the 800 human miRNAs Nanostring panel. Nanostring nCounter Analysis System generated data, then Partek GS software was used for secondary analysis. RESULTS Cytori processing resulted in a higher cell yield at isolation (p= 0.0156), and ADSCs exhibited higher proliferation in vitro at P0 (n. of days at P0, p=0.0313), higher cell yield at P1 (p=0.0313), and higher PDL1 (p=0.0469). Out of the 361 markers examined, in every donor at each passage, at least 94 were found positive (expression >1%), including 30 within the newly screened 117 markers. Altogether, 27 markers were expressed at more than 85%, including 6 from the newly screened markers. Our analysis identified 50 donor-dependent markers, 50 passage-modulated markers (P2 vs P3) and 45 IFN-g-modulated marker, 41 increased and 4 decreased. We were able to identify the ADSCs progenitors markers CD146 and CD271 on cell as in P0 and P3, we observed a transition from a dominant expression in P0 of CD271 (34.05%) vs CD146 (1.62% ) to a dominant expression of CD146 in P3 (52%) vs CD271 (0.7%) in a single donor. The frequency of double positives CD146+CD271+ was 1% and 0.6% in P0 and P3 respectively. miRNA expression analysis by ANOVA revealed that (i) IFN-g-priming modulated 19 miRNAs, that 4 miRNAs were modulated by passage number and donors' origin. CONCLUSION Our study shows that the standardized Cytori processing advantageously substitutes AT manual digestion by enabling higher cell yield and potentially providing multiple ADSC doses from a single donor. The isolation procedure is standardized, the operator-dependent variations are minimized, and less prone to contamination compared to the lengthy, multistep process of manual digestion. The broad cell characterization with flow cytometry and miRNA expression revealed the expression and modulation of new markers. We believe that increasing the dimensionality of the ADSC characterization, beyond the traditional markers, could reveal markers that describe specific functional abilities of clinical ADSC product. Proper functional studies are necessary to validate our hypothesis. Disclosures No relevant conflicts of interest to declare.

Endogenous Wnt Signaling Promotes Proliferation and Suppresses Osteogenic Differentiation in Human Adipose Derived Stromal Cells

2006 ◽  
Vol 0 (0) ◽  
pp. 060210065733001
Author(s):  
Hyun Hwa Cho ◽  
Yeon Jeong Kim ◽  
Su Jin Kim ◽  
Jae Ho Kim ◽  
Yong Chan Bae ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Adipose Derived Stromal Cells

scholarly journals Adipose-Derived Stromal Cells Seeded on Integra® Dermal Regeneration Template Improve Post-Burn Wound Reconstruction

2020 ◽  
Vol 7 (3) ◽  
pp. 67
Author(s):  
Marcin Piejko ◽  
Karolina Radziun ◽  
Sylwia Bobis-Wozowicz ◽  
Agnieszka Waligórska ◽  
Eliza Zimoląg ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Experimental Treatment ◽  
Collagenolytic Activity ◽  
Burn Wound ◽  
Clinical Issue ◽  
Excision Wound ◽  
Adipose Derived Stromal Cells ◽  
Dermal Regeneration Template ◽  
Endothelial Growth Factor

Fibrosis of burn-related wounds remains an unresolved clinical issue that leads to patient disability. The aim of this study was to assess the efficacy of the transplantation of adipose-derived stromal cells seeded onto a collagen-based matrix in the reconstruction of burn-related scars. Here, we characterized an in vitro interaction between adipose-derived stromal cells and a collagen-based matrix, Integra®DRT. Our results show that transcription of pro-angiogenic, remodeling, and immunomodulatory factors was more significant in adipose-derived stromal cells than in fibroblasts. Transcription of metalloproteinases 2 and 9 is positively correlated with the collagenolytic activity of the adipose-derived stromal cells seeded onto Integra®DRT. The increase in the enzymatic activity corresponds to the decrease in the elasticity of the whole construct. Finally, we validated the treatment of a post-excision wound using adipose-derived stromal cells and an Integra®DRT construct in a 25-year-old woman suffering from burn-related scars. Scarless healing was observed in the area treated by adipose-derived stromal cells and the Integra®DRT construct but not in the reference area where Integra®DRT was applied without cells. This clinical observation may be explained by in vitro findings: Enhanced transcription of the vascular endothelial growth factor as well as remodeling of the collagen-based matrix decreased mechanical stress. Our experimental treatment demonstrated that the adipose-derived stromal cells seeded onto Integra®DRT exhibit valuable properties that may improve post-excision wound healing and facilitate skin regeneration without scars.

Cartilage Particles can Promote Chondrogenesis of Adipose-Derived Stromal Cells on Poly(ε-Caprolactone)/Fibrin Hybrid Constructs Prepared via Sandwich Model

2020 ◽  
Vol 47 ◽  
pp. 63-74
Author(s):  
Sahar Ghosouri ◽  
Mohsen Setayeshmehr ◽  
Asghar Taheri-Kafrani ◽  
Ali Valiani
Keyword(s):  
Stromal Cells ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Layer By Layer ◽  
Cell Seeding ◽  
Rt Pcr ◽  
Electrospun Scaffolds ◽  
Adipose Derived Stromal Cells ◽  
Histological Results

Electrospun fibers have demonstrated a remarkable potential as a framework structure in the fabrication of cartilage tissue engineering (CTE) scaffolds. Various extracellular matrices have been incorporated into electrospun scaffolds to mimic and simulate the extracellular environment. The objective of this study was to fabricate hybrid constructs using composite electrospun scaffolds based on poly (ε-caprolactone) (PCL) and cartilage-derived matrix (CDM) and fibrin hydrogel to improve the viability and differentiation of human adipose-derived stromal cells (ADSCs) for CTE applications.Initially, PCL and PCL-CDM electrospun mats were fabricated. Fibrin/ ADSCs hydrogel were seeded on PCL- CDM mats and arranged layer-by-layer using sandwich technique. This method has been employed to increase cell seeding and infiltration efficiency through the entire mass of the scaffold. Real-time reverse-transcription polymerase chain reaction (RT- PCR), were performed to examine the expression of collagen types II and X, SOX9 and aggrecan. The production of glycosaminoglycan (GAG) was also tested in vitro by Toluidine blue stain and biochemical assay in the cultured scaffolds.The findings demonstrated that incorporation of CDM in PCL fibers results in improved cell viability. Hematoxylin and eosin staining showed that the sandwich method resulted in homogenous cell seeding within the scaffold. Overall, the RT- PCR, biochemical and histological results, showed that incorporation of the CDM into PCL/fibrin sandwich scaffolds stimulated ADSCs chondrogenesis and produced the products which increased expression of chondrogenic genes. It also, enhanced GAG synthesis compared to PCL/fibrin scaffolds.These findings suggest PCL-CDM/fibrin can be considered as an appropriate hybrid scaffold for CTE applications.

Sign in / Sign up

Export Citation Format

Share Document