scholarly journals Modulation of Human Adipose Stem Cells’ Neurotrophic Capacity Using a Variety of Growth Factors for Neural Tissue Engineering Applications: Axonal Growth, Transcriptional, and Phosphoproteomic Analyses In Vitro

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1939
Author(s):  
Katharina M. Prautsch ◽  
Alexander Schmidt ◽  
Viola Paradiso ◽  
Dirk J. Schaefer ◽  
Raphael Guzman ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Neurotrophic Factor ◽  
Axonal Growth ◽  
Cellular Level ◽  
Adipose Stem Cells ◽  
Transcriptional Analysis ◽  
Axonal Outgrowth ◽  
Human Adipose Stem Cells

We report on a potential strategy involving the exogenous neurotrophic factors (NTF) for enhancing the neurotrophic capacity of human adipose stem cells (ASC) in vitro. For this, ASC were stimulated for three days using NTF, i.e., nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT3), NT4, glial cell-derived neurotrophic factor (GDNF), and ciliary neurotrophic factor (CNTF). The resulting conditioned medium (CM) as well as individual NTF exhibited distinct effects on axonal outgrowth from dorsal root ganglion (DRG) explants. In particular, CM derived from NT3-stimulated ASC (CM-NT3-ASC) promoted robust axonal outgrowth. Subsequent transcriptional analysis of DRG cultures in response to CM-NT3-ASC displayed significant upregulation of STAT-3 and GAP-43. In addition, phosphoproteomic analysis of NT3-stimulated ASC revealed significant changes in the phosphorylation state of different proteins that are involved in cytokine release, growth factors signaling, stem cell maintenance, and differentiation. Furthermore, DRG cultures treated with CM-NT3-ASC exhibited significant changes in the phosphorylation levels of proteins involved in tubulin and actin cytoskeletal pathways, which are crucial for axonal growth and elongation. Thus, the results obtained at the transcriptional, proteomic, and cellular level reveal significant changes in the neurotrophic capacity of ASC following NT3 stimulation and provide new options for improving the axonal growth-promoting potential of ASC in vitro.

scholarly journals Effects of different serum conditions on osteogenic differentiation of human adipose stem cells in vitro

2013 ◽  
Vol 4 (1) ◽  
pp. 17 ◽  
Author(s):  
Laura Kyllönen ◽  
Suvi Haimi ◽  
Bettina Mannerström ◽  
Heini Huhtala ◽  
Kristiina M Rajala ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Adipose Stem Cells ◽  
Human Adipose Stem Cells

scholarly journals Ex-Vivo Stimulation of Adipose Stem Cells by Growth Factors and Fibrin-Hydrogel Assisted Delivery Strategies for Treating Nerve Gap-Injuries

2020 ◽  
Vol 7 (2) ◽  
pp. 42
Author(s):  
Katharina M. Prautsch ◽  
Lucas Degrugillier ◽  
Dirk J. Schaefer ◽  
Raphael Guzman ◽  
Daniel F. Kalbermatten ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Growth Factor ◽  
Nerve Regeneration ◽  
Ex Vivo ◽  
Axonal Outgrowth ◽  
Assisted Delivery ◽  
Delivery Strategies ◽  
Stimulation Of

Peripheral nerve injuries often result in lifelong disabilities despite advanced surgical interventions, indicating the urgent clinical need for effective therapies. In order to improve the potency of adipose-derived stem cells (ASC) for nerve regeneration, the present study focused primarily on ex-vivo stimulation of ASC by using growth factors, i.e., nerve growth factor (NGF) or vascular endothelial growth factor (VEGF) and secondly on fibrin-hydrogel nerve conduits (FNC) assisted ASC delivery strategies, i.e., intramural vs. intraluminal loading. ASC were stimulated by NGF or VEGF for 3 days and the resulting secretome was subsequently evaluated in an in vitro axonal outgrowth assay. For the animal study, a 10 mm sciatic nerve gap-injury was created in rats and reconstructed using FNC loaded with ASC. Secretome derived from NGF-stimulated ASC promoted significant axonal outgrowth from the DRG-explants in comparison to all other conditions. Thus, NGF-stimulated ASC were further investigated in animals and found to enhance early nerve regeneration as evidenced by the increased number of β-Tubulin III+ axons. Notably, FNC assisted intramural delivery enabled the improvement of ASC’s therapeutic efficacy in comparison to the intraluminal delivery system. Thus, ex-vivo stimulation of ASC by NGF and FNC assisted intramural delivery may offer new options for developing effective therapies.

Buccal Fat Pad, an Oral Access Source of Human Adipose Stem Cells with Potential for Osteochondral Tissue Engineering: An In Vitro Study

2010 ◽  
Vol 16 (5) ◽  
pp. 1083-1094 ◽  
Author(s):  
Elisabet Farré-Guasch ◽  
Carles Martí-Pagès ◽  
Federico Hernández-Alfaro ◽  
Jenneke Klein-Nulend ◽  
Núria Casals
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
In Vitro Study ◽  
Adipose Stem Cells ◽  
Vitro Study ◽  
Fat Pad ◽  
Buccal Fat Pad ◽  
Osteochondral Tissue Engineering ◽  
Human Adipose Stem Cells

scholarly journals Enhanced Osteogenic and Vasculogenic Differentiation Potential of Human Adipose Stem Cells on Biphasic Calcium Phosphate Scaffolds in Fibrin Gels

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Fransisca A. S. van Esterik ◽  
Behrouz Zandieh-Doulabi ◽  
Cornelis J. Kleverlaan ◽  
Jenneke Klein-Nulend
Keyword(s):  
Stem Cells ◽  
Calcium Phosphate ◽  
Biphasic Calcium Phosphate ◽  
Adipose Stem Cells ◽  
Differentiation Potential ◽  
Fibrin Gels ◽  
Enhanced Expression ◽  
Human Adipose Stem Cells

For bone tissue engineering synthetic biphasic calcium phosphate (BCP) with a hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ratio of 60/40 (BCP60/40) is successfully clinically applied, but the high percentage of HA may hamper efficient scaffold remodelling. Whether BCP with a lower HA/β-TCP ratio (BCP20/80) is more desirable is still unclear. Vascular development is needed before osteogenesis can occur. We aimed to test the osteogenic and/or vasculogenic differentiation potential as well as degradation of composites consisting of human adipose stem cells (ASCs) seeded on BCP60/40 or BCP20/80 incorporated in fibrin gels that trigger neovascularization for bone regeneration. ASC attachment to BCP60/40 and BCP20/80 within 30 min was similar (>93%). After 11 days of culture BCP20/80-based composites showed increased alkaline phosphatase activity andDMP1gene expression, but notRUNX2and osteonectin expression, compared to BCP60/40-based composites. BCP20/80-based composites also showed enhanced expression of the vasculogenic markersCD31andVEGF189, but notVEGF165and endothelin-1. Collagen-1 and collagen-3 expression was similar in both composites. Fibrin degradation was increased in BCP20/80-based composites at day 7. In conclusion, BCP20/80-based composites showed enhanced osteogenic and vasculogenic differentiation potential compared to BCP60/40-based compositesin vitro, suggesting that BCP20/80-based composites might be more promising forin vivobone augmentation than BCP60/40-based composites.

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