scholarly journals The Development And Characterization Of Heparin-Conjugated Fibrin Hydrogel With Incapsulated Mesenchimal Stem Cells And Growth Factors

2021 ◽  
Author(s):  
M.A. Sarsenova ◽  
A.S. Issabekova ◽  
M.R. Karzhauov ◽  
G.K. Kudaibergen ◽  
M.S. Zhunussova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Regenerative Medicine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Stem Cells ◽  
Carbodiimide Method ◽  
Kinetics Of ◽  
Tgf Β1

Currently, one of the major focuses in regenerative medicine is the development and implementation into practice of composite biomaterials with chondro- and osteoinductive properties, which include human stem cells and growth factors. Heparin-conjugated fibrinogen was obtained using the carbodiimide method, which was further used to create heparin-conjugated fibrin hydrogels (HCFH). As a result of this work, two types of HCFH were obtained: a hydrogel with encapsulated mesenchymal stem cells (MSC) and a hydrogel with TGF-β1 and BMP-4 growth factors. It has been found that synovial MSCs retain viability after encapsulation in HCFH, which indicates that the developed hydrogel is biocompatible and does not have toxic effect to the cells. The results of enzyme-linked immunosorbent assay on the kinetics of BMP-4 and TGF-β1 release from HCFH showed that the developed hydrogel is able to retain BMP-4 and TGF-β1. The kinetics of release from HCFH into phosphate buffer was significantly slower compared to fibrin hydrogel.

scholarly journals Concise Review: Multifaceted Characterization of Human Mesenchymal Stem Cells for Use in Regenerative Medicine

2017 ◽  
Vol 6 (12) ◽  
pp. 2173-2185 ◽  
Author(s):  
Rebekah M. Samsonraj ◽  
Michael Raghunath ◽  
Victor Nurcombe ◽  
James H. Hui ◽  
Andre J. van Wijnen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Human Mesenchymal Stem Cells ◽  
Concise Review

scholarly journals A Simplified Method for the Aspiration of Bone Marrow from Patients Undergoing Hip and Knee Joint Replacement for Isolating Mesenchymal Stem Cells and In Vitro Chondrogenesis

2016 ◽  
Vol 2016 ◽  
pp. 1-18 ◽  
Author(s):  
Subhash C. Juneja ◽  
Sowmya Viswanathan ◽  
Milan Ganguly ◽  
Christian Veillette
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Isolation And Characterization ◽  
Standard Operating ◽  
Total Knee ◽  
Detailed Protocol

The procedure for aspiration of bone marrow from the femur of patients undergoing total knee arthroplasty (TKA) or total hip arthroplasty (THA) may vary from an OR (operating room) to OR based on the surgeon’s skill and may lead to varied extent of clotting of the marrow and this, in turn, presents difficulty in the isolation of mesenchymal stem cells (MSCs) from such clotted bone marrow. We present a simple detailed protocol for aspirating bone marrow from such patients, isolation, and characterization of MSCs from the aspirated bone marrow specimens and show that the bone marrow presented no clotting or exhibited minimal clotting. This represents an economical source and convenient source of MSCs from bone marrow for use in regenerative medicine. Also, we presented the detailed protocol and showed that the MSCs derived from such bone marrow specimens exhibited MSCs characteristics and generated micromass cartilages, the recipe for regenerative medicine for osteoarthritis. The protocols we presented can be used as standard operating procedures (SOPs) by researchers and clinicians.

scholarly journals Mesenchymal Stem Cells Desensitize Castration-resistant Prostate Cancer to Docetaxel Chemotherapy via Inducing TGF-β1-mediated Cell Autophagy

2020 ◽  
Author(s):  
Yang Yu ◽  
Wen-tao Zhang ◽  
Fu-han Yang ◽  
Ya-dong Guo ◽  
Lin Ye ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Castration Resistant Prostate Cancer ◽  
Castration Resistant ◽  
Docetaxel Chemotherapy ◽  
Tgf Β1

Abstract Background: Mesenchymal stem cells (MSCs) have been proved to accelerate prostate cancer (PCa) castration resistance progression. The purpose of this study is to investigate the contribution of MSCs to the development of docetaxel resistance in castration-resistant prostate cancer (CRPC) cells and its potential mechanisms.Methods: The effect of MSCs on CRPC cells resistance to docetaxel was determined using in-vivo and in-vitro approaches. CCK8 and PI/Annexin V-FITC assay were used to examined the cell viability and apoptosis. The concentration of transforming growth factor-β1 was measured by enzyme-linked immunosorbent assay and small interfering RNA was used for functional analyses.Results: MSCs significantly reduced the sensitivity of CRPC cells to docetaxel-induced proliferation inhibition and apoptosis promotion in vivo and in vitro. CRPC cells cocultured with MSCs under docetaxel administration have an increased autophagy activation, while autophagy inhibitor could effectively reversed MSCs-induced resistance to docetaxel. Additionally, MSCs-induced CRPC cell autophagy increase under docetaxel administration depends on MSCs secreting TGF-β1 and inhibition of TGF-β1 secretion in MSCs could consequently increase the sensitivity of CRPC cells to docetaxel.Conclusions: These results suggest that docetaxel administrated CRPC cells may elicit MSCs secreting TGF-β1 increase, which desensitizes CRPC to docetaxel chemotherapy accelerating chemoresistance occurrence via inducing cell autophagy.

АДАПТАЦИЯ МЕТОДА ИНТЕРФЕРОМЕТРИИ СЛОЯ БИОМОЛЕКУЛ ДЛЯ КОЛИЧЕСТВЕННОЙ ОЦЕНКИ СОДЕРЖАНИЯ ФАКТОРА РОСТА ЭНДОТЕЛИЯ СОСУДОВ В КОНДИЦИОНИРОВАННОЙ КЛЕТОЧНОЙ СРЕДЕ

2020 ◽  
Vol 65 (6) ◽  
pp. 1099-1106
Author(s):  
М.В. Волкова ◽  
◽  
В.В. Бояринцев ◽  
А.В. Трофименко ◽  
С.А. Бирюков ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
High Sensitivity ◽  
Conditioned Media ◽  
Enzyme Linked Immunosorbent Assay ◽  
Growth Media ◽  
Cell Sheets ◽  
Hypoxic Conditions ◽  
Endothelial Growth Factor

Quantitative analysis of cytokines, chemokines, growth factors and other soluble proteins in different biological liquids is routinely performed in contemporary diagnostics and biomedicine research. However, current methods of analysis are time-consuming and include multiple steps. In this study, we have developed a protocol that describes how bio-layer interferometry can be applied to quantify an analyte in several minutes. Conditioned growth media collected from mouse mesenchymal stem cells grown in normoxia or hypoxic conditions in a monolayer fashion, MSC-derived 3D cell sheets and 3D spheroids were used as a model system in which we determined a concentration of vascular endothelial growth factor (VEGF-A). This technique displayed a high sensitivity (down to 0.1 ng of VEGF-A per mL as a minimum). The measured concentrations of VEGF-A in the conditioned media from mesenchymal stem cells turned out to be similar with values determined by the enzyme-linked immunosorbent assay. Using bio-layer interferometry, it was shown that as compared to mesenchymal stem cells grown in monolayer, spheroids and 3D sheets of mesenchymal stem cells produce significantly more VEGF-A (by 2.5-3.0-fold). Thus, due to the developed protocol it was possible to adapt bio-layer interferometry for rapid quantification of growth factors in conditioned media.

scholarly journals Optimizing growth factor induction of tenogenesis in three-dimensional culture of mesenchymal stem cells

2019 ◽  
Vol 10 ◽  
pp. 204173141984877 ◽  
Author(s):  
Ibtesam Rajpar ◽  
Jennifer G Barrett
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Growth Factors ◽  
Tendon Healing ◽  
Collagen Type I ◽  
Marker Genes ◽  
Type I ◽  
Cell Alignment ◽  
Tgf Β1

Adult tissue stem cells have shown promise for the treatment of debilitating tendon injuries. However, few comparisons of stem cells from different tissue sources have been made to determine the optimum stem cell source for treating tendon. Moreover, it is likely that the application of tenogenic growth factors will improve tendon stem cell treatments further, and a comprehensive comparison of a number of growth factors is needed. Thus far, different types of stem cells cannot be evaluated in a high-throughput manner. To this end, we have developed an approach to culture mesenchymal stem cells isolated from bone marrow in collagen type I hydrogels with tenogenic growth factors using economical, commercially available supplies. To optimize growth factors for this assay, FGF-2, TGF-β1, IGF-1, and/or BMP-12 were tested singly and in novel combinations of (1) BMP-12 and IGF-1, (2) TGF-β1 and IGF-1, and/or (3) BMP-12 and FGF-2 over 10 days. Our data suggest that BMP-12 supplementation alone results in the strongest expression of tendon marker genes, controlled contractility of constructs, a higher degree of cell alignment, and tendon-like tissue morphology. This easy-to-use benchtop assay can be used to screen novel sources of stem cells and cell lines for tissue engineering and tendon healing applications.

scholarly journals Mesenchymal stem cells desensitize castration-resistant prostate cancer to docetaxel chemotherapy via inducing TGF-β1-mediated cell autophagy

2020 ◽  
Author(s):  
Yang Yu ◽  
Wen-tao Zhang ◽  
Fu-han Yang ◽  
Ya-dong Guo ◽  
Lin Ye ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Castration Resistant Prostate Cancer ◽  
Castration Resistant ◽  
Docetaxel Chemotherapy ◽  
Tgf Β1

Abstract Background: Mesenchymal stem cells (MSCs) have been proved to accelerate prostate cancer (PCa) castration resistance progression. The purpose of this study is to investigate the contribution of MSCs to the development of docetaxel resistance in castration-resistant prostate cancer (CRPC) cells and its potential mechanisms. Methods: The effect of MSCs on CRPC cells resistance to docetaxel was determined using in-vivo and in-vitro approaches. CCK8 and PI/Annexin V-FITC assay were used to examined the cell viability and apoptosis. The concentration of transforming growth factor-β1 was measured by enzyme-linked immunosorbent assay and small interfering RNA was used for functional analyses. Results: MSCs significantly reduced the sensitivity of CRPC cells to docetaxel-induced proliferation inhibition and apoptosis promotion in vivo and in vitro. CRPC cells cocultured with MSCs under docetaxel administration have an increased autophagy activation, while autophagy inhibitor could effectively reversed MSCs-induced resistance to docetaxel. Additionally, MSCs-induced CRPC cell autophagy increase under docetaxel administration depends on MSCs secreting TGF-β1 and inhibition of TGF-β1 secretion in MSCs could consequently increase the sensitivity of CRPC cells to docetaxel. Conclusions: These results suggest that docetaxel administrated CRPC cells may elicit MSCs secreting TGF-β1 increase, which desensitizes CRPC to docetaxel chemotherapy accelerating chemoresistance occurrence via inducing cell autophagy.

scholarly journals Mesenchymal stem cells promote pancreatic β-cell regeneration through downregulation of FoxO1 pathway

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Rahul Khatri ◽  
Sybille Mazurek ◽  
Sebastian Friedrich Petry ◽  
Thomas Linn
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Proinflammatory Cytokines ◽  
Enzyme Linked Immunosorbent Assay ◽  
Intravenous Route ◽  
Β Cells ◽  
Β Cell ◽  
Pancreatic Β Cells ◽  
Alu Sequence

Abstract Background Mesenchymal stem cells (MSC) are non-haematopoietic, fibroblast-like multipotent stromal cells. In the injured pancreas, these cells are assumed to secrete growth factors and immunomodulatory molecules, which facilitate the regeneration of pre-existing β-cells. However, when MSC are delivered intravenously, their majority is entrapped in the lungs and does not reach the pancreas. Therefore, the aim of this investigation was to compare the regenerative support of hTERT-MSC (human telomerase reverse transcriptase mesenchymal stem cells) via intrapancreatic (IPR) and intravenous route (IVR). Methods hTERT-MSC were administered by IPR and IVR to 50% pancreatectomized NMRI nude mice. After eight days, blood glucose level, body weight, and residual pancreatic weight were measured. Proliferating pancreatic β-cells were labelled and identified with bromodeoxyuridine (BrdU) in vivo. The number of residual islets and the frequency of proliferating β-cells were compared in different groups with sequential pancreatic sections. The pancreatic insulin content was evaluated by enzyme-linked immunosorbent assay (ELISA) and the presence of hTERT-MSC with human Alu sequence. Murine gene expression of growth factors, β-cell specific molecules and proinflammatory cytokines were inspected by real-time polymerase chain reaction (RT-PCR) and Western blot. Results This study evaluated the regenerative potential of the murine pancreas post-hTERT-MSC administration through the intrapancreatic (IPR) and intravenous route (IVR). Both routes of hTERT-MSC transplantation (IVR and IPR) increased the incorporation of BrdU by pancreatic β-cells compared to control. MSC induced epidermal growth factor (EGF) expression and inhibited proinflammatory cytokines (IFN-γ and TNF-α). FOXA2 and PDX-1 characteristics for pancreatic progenitor cells were activated via AKT/ PDX-1/ FoxO1 signalling pathway. Conclusion The infusion of hTERT-MSC after partial pancreatectomy (Px) through the IVR and IPR facilitated the proliferation of autochthonous pancreatic β-cells and provided evidence for a regenerative influence of MSC on the endocrine pancreas. Moderate benefit of IPR over IVR was observed which could be a new treatment option for preventing diabetes mellitus after pancreas surgery.

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