Critical steps in the isolation and expansion of adipose-derived stem cells for translational therapy

2015 ◽  
Vol 17 ◽  
Author(s):  
S. Riis ◽  
V. Zachar ◽  
S. Boucher ◽  
M.C. Vemuri ◽  
C. P. Pennisi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Standard Treatment ◽  
Treatment Options ◽  
Good Manufacturing Practice ◽  
European Medicines Agency ◽  
Adipose Derived Stem Cells ◽  
High Expectations ◽  
Beneficial Effects ◽  
Expansion Protocol ◽  
Medical Effectiveness

Since the discovery of adipose-derived stem cells (ASCs), there have been high expectations of their putative clinical use. Recent advances support these expectations, and it is expected that the transition from pre-clinical and clinical studies to implementation as a standard treatment modality is imminent. However ASCs must be isolated and expanded according to good manufacturing practice guidelines and a basic assurance of quality, safety, and medical effectiveness is needed for authorisation by regulatory agencies, such as European Medicines Agency and US Food and Drug Administration. In this review, a collection of studies investigating the influence of different steps of the isolation and expansion protocol on the yield and functionality of ASCs has been presented in an attempt to come up with best recommendations that ensure potential beneficial clinical outcome of using ASCs in any therapeutic setting. If the findings confirm the initial observations of beneficial effects of ASCs, the path is paved for implementing these ASC-based therapies as standard treatment options.

scholarly journals The Crosstalk of Adipose-Derived Stem Cells (ADSC), Oxidative Stress, and Inflammation in Protective and Adaptive Responses

2020 ◽  
Vol 21 (23) ◽  
pp. 9262
Author(s):  
Luc Rochette ◽  
Loubna Mazini ◽  
Gabriel Malka ◽  
Marianne Zeller ◽  
Yves Cottin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Intracellular Signaling ◽  
Adipose Derived Stem Cells ◽  
Adaptive Responses ◽  
Complex Signals ◽  
Beneficial Effects ◽  
Secondary Messenger ◽  
Secreted Factors ◽  
Oxidation Reduction ◽  
Therapeutic Properties

The potential use of stem cell-based therapies for the repair and regeneration of various tissues and organs is a major goal in repair medicine. Stem cells are classified by their potential to differentiate into functional cells. Compared with other sources, adipose-derived stem cells (ADSCs) have the advantage of being abundant and easy to obtain. ADSCs are considered to be tools for replacing, repairing, and regenerating dead or damaged cells. The capacity of ADSCs to maintain their properties depends on the balance of complex signals in their microenvironment. Their properties and the associated outcomes are in part regulated by reactive oxygen species, which mediate the oxidation-reduction state of cells as a secondary messenger. ADSC therapy has demonstrated beneficial effects, suggesting that secreted factors may provide protection. There is evidence that ADSCs secrete a number of cytokines, growth factors, and antioxidant factors into their microenvironment, thus regulating intracellular signaling pathways in neighboring cells. In this review, we introduce the roles of ADSCs in the protection of cells by modulating inflammation and immunity, and we develop their potential therapeutic properties.

scholarly journals Effect of Nanostructured Scaffold on Human Adipose-Derived Stem Cells: Outcome of In Vitro Experiments

Nanomaterials ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1822 ◽  
Author(s):  
Marina Borgese ◽  
Ludovica Barone ◽  
Federica Rossi ◽  
Mario Raspanti ◽  
Roberto Papait ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Medium ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adipose Derived Stem Cells ◽  
In Vitro Experiments ◽  
Fatty Acid Binding ◽  
Beneficial Effects ◽  
Angiogenesis Process

This work is addressed to provide, by in vitro experiments, results on the repercussion that a nanostructured scaffold could have on viability, differentiation and secretion of bioactive factors of human adipose-derived stem cells (hASCs) when used in association to promote angiogenesis, a crucial condition to favour tissue regeneration. To achieve this aim, we evaluated cell viability and morphology by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and microscopy analysis, respectively. We also investigated the expression of some of those genes involved in angiogenesis and differentiation processes utilizing quantitative polymerase chain reaction (qPCR), whereas the amounts of Vascular Endothelial Growth Factor A, Interleukin 6 and Fatty Acid-Binding Protein 4 secreted in the culture medium, were quantified by enzyme-linked immunosorbent assay (ELISA). Results suggested that, in the presence of the scaffold, cell proliferation and the exocytosis of factors involved in the angiogenesis process are reduced; by contrast, the expression of those genes involved in hASC differentiation appeared enhanced. To guarantee cell survival, the construct dimensions are, generally, smaller than clinically required. Furthermore, being the paracrine event the primary mechanism exerting the beneficial effects on injured tissues, the use of conditioned culture medium instead of cells may be convenient.

scholarly journals Thymosin β4-Enhancing Therapeutic Efficacy of Human Adipose-Derived Stem Cells in Mouse Ischemic Hindlimb Model

2020 ◽  
Vol 21 (6) ◽  
pp. 2166
Author(s):  
Jong-Ho Kim ◽  
I-Rang Lim ◽  
Chi-Yeon Park ◽  
Hyung Joon Joo ◽  
Ji-Min Noh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Intracellular Signaling ◽  
Morphological Changes ◽  
Additional Treatment ◽  
Endothelial Differentiation ◽  
Adipose Derived Stem Cells ◽  
Thymosin Β4 ◽  
Differentiation Potential ◽  
Beneficial Effects

Thymosin β4 (Tβ4) is a G-actin sequestering protein that contributes to diverse cellular activities, such as migration and angiogenesis. In this study, the beneficial effects of combined cell therapy with Tβ4 and human adipose-derived stem cells (hASCs) in a mouse ischemic hindlimb model were investigated. We observed that exogenous treatment with Tβ4 enhanced endogenous TMSB4X mRNA expression and promoted morphological changes (increased cell length) in hASCs. Interestingly, Tβ4 induced the active state of hASCs by up-regulating intracellular signaling pathways including the PI3K/AKT/mTOR and MAPK/ERK pathways. Treatment with Tβ4 significantly increased cell migration and sprouting from microbeads. Moreover, additional treatment with Tβ4 promoted the endothelial differentiation potential of hASCs by up-regulating various angiogenic genes. To evaluate the in vivo effects of the Tβ4-hASCs combination on vessel recruitment, dorsal window chambers were transplanted, and the co-treated mice were found to have a significantly increased number of microvessel branches. Transplantation of hASCs in combination with Tβ4 was found to improve blood flow and attenuate limb or foot loss post-ischemia compared to transplantation with hASCs alone. Taken together, the therapeutic application of hASCs combined with Tβ4 could be effective in enhancing endothelial differentiation and vascularization for treating hindlimb ischemia.

Good manufacturing practice-compliant isolation and culture of human adipose derived stem cells

2014 ◽  
Vol 1 (4) ◽  
Author(s):  
Phuc Van Pham ◽  
Ngoc Bich Vu ◽  
Nhan Lu-Chinh Phan ◽  
Dung Minh Le ◽  
Nhat Chau Truong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Good Manufacturing Practice ◽  
Adipose Derived Stem Cells

scholarly journals 3D Bioprinting of Human Adipose-Derived Stem Cells and Their Tenogenic Differentiation in Clinical-Grade Medium

2020 ◽  
Vol 21 (22) ◽  
pp. 8694
Author(s):  
Deborah Stanco ◽  
Monica Boffito ◽  
Alessia Bogni ◽  
Luca Puricelli ◽  
Josefa Barrero ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Viability ◽  
Scale Up ◽  
Good Manufacturing Practice ◽  
Adipose Derived Stem Cells ◽  
3D Bioprinting ◽  
Clinical Scale ◽  
Nanofibrillar Cellulose ◽  
Tenogenic Differentiation ◽  
Tendon Tissue Engineering

Defining the best combination of cells and biomaterials is a key challenge for the development of tendon tissue engineering (TE) strategies. Adipose-derived stem cells (ASCs) are ideal candidates for this purpose. In addition, controlled cell-based products adherent to good manufacturing practice (GMP) are required for their clinical scale-up. With this aim, in this study, ASC 3D bioprinting and GMP-compliant tenogenic differentiation were investigated. In detail, primary human ASCs were embedded within a nanofibrillar-cellulose/alginate bioink and 3D-bioprinted into multi-layered square-grid matrices. Bioink viscoelastic properties and scaffold ultrastructural morphology were analyzed by rheology and scanning electron microscopy (SEM). The optimal cell concentration for printing among 3, 6 and 9 × 106 ASC/mL was evaluated in terms of cell viability. ASC morphology was characterized by SEM and F-actin immunostaining. Tenogenic differentiation ability was then evaluated in terms of cell viability, morphology and expression of scleraxis and collagen type III by biochemical induction using BMP-12, TGF-β3, CTGF and ascorbic acid supplementation (TENO). Pro-inflammatory cytokine release was also assessed. Bioprinted ASCs showed high viability and survival and exhibited a tenocyte-like phenotype after biochemical induction, with no inflammatory response to the bioink. In conclusion, we report a first proof of concept for the clinical scale-up of ASC 3D bioprinting for tendon TE.

scholarly journals Intramyocardial injection of human adipose-derived stem cells ameliorates cognitive deficit by regulating oxidative stress–mediated hippocampal damage after myocardial infarction

2021 ◽  
Author(s):  
Tsung-Ming LEE ◽  
Cheng-Che LEE ◽  
Horng-Jyh HARN ◽  
Tzyy-Wen Chiou ◽  
Ming-Hsi CHUANG ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cells ◽  
Cognitive Impairment ◽  
Cognitive Deficit ◽  
Adipose Derived Stem Cells ◽  
Oxygen Species ◽  
Intramyocardial Injection ◽  
Beneficial Effects ◽  
Therapeutic Benefits ◽  
Cell Engraftment

Abstract Cognitive impairment is a serious side effect of post-myocardial infarction (MI) course. We have recently demonstrated that human adipose-derived stem cells (hADSCs) ameliorated myocardial injury after MI by attenuating reactive oxygen species (ROS) levels. Here, we studied whether the beneficial effects of intramyocardial hADSC transplantation can extend to the brain and how they may attenuate cognitive dysfunction via modulating ROS after MI. After coronary ligation, male Wistar rats were randomized via an intramyocardial route to receive either vehicle, hADSC transplantation (1 × 106 cells), or the combination of hADSCs and 3-Morpholinosydnonimine (SIN-1, a peroxynitrite donor). Whether hADSCs migrated into the hippocampus was assessed by using human-specific primers in qPCR reactions. Passive avoidance test was used to assess cognitive performance. Postinfarction was associated with increased oxidative stress in the myocardium, circulation, and hippocampus. This was coupled with decreased numbers of dendritic spines as well as a significant downregulation of synaptic plasticity consisting of synaptophysin and PSD95. Step-through latency during passive avoidance test was impaired in vehicle-treated rats after MI. Intramyocardial hADSC injection exerted therapeutic benefits in improving cardiac function and cognitive impairment. None of hADSCs was detected in rat’s hippocampus at the 3rd day after intramyocardial injection. The beneficial effects of hADSCs on MI-induced histological and cognitive changes were abolished after adding SIN-1. MI-induced ROS attacked the hippocampus to induce neurodegeneration, resulting in cognitive deficit. The remotely intramyocardial administration of hADSCs has the capacity of improved synaptic neuroplasticity in the hippocampus mediated by ROS, not the cell engraftment, after MI. Key messages Human adipose-derived stem cells (hADSCs) ameliorated injury after myocardial infarction by attenuating reactive oxygen species (ROS) levels. Intramyocardial administration of hADSCs remotely exerted therapeutic benefits in improving cognitive impairment after myocardial infarction. The improved synaptic neuroplasticity in the hippocampus was mediated by hADSC-inhibiting ROS, not by the stem cell engraftment.

scholarly journals Intramyocardial Injection of Human Adipose-Derived Stem Cells Ameliorates Cognitive Deficit By Regulating Oxidative Stress-Mediated Hippocampal Damage After Myocardial Infarction

2021 ◽  
Author(s):  
Tsung-Ming Lee ◽  
Cheng-Che LEE ◽  
Horng-Jyh HARN ◽  
Tzyy-Wen CHIOU ◽  
Ming-Hsi CHUANG ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Myocardial Infarction ◽  
Stem Cells ◽  
Cognitive Impairment ◽  
Passive Avoidance ◽  
Cognitive Deficit ◽  
Passive Avoidance Test ◽  
Adipose Derived Stem Cells ◽  
Intramyocardial Injection ◽  
Beneficial Effects

Abstract Cognitive impairment is a serious side effect of post-myocardial infarction (MI) course. We have recently demonstrated that human adipose-derived stem cells (hADSCs) ameliorated myocardial injury after MI by attenuating reactive oxygen species (ROS) levels. Here, we studied whether the beneficial effects of intramyocardial hADSC transplantation can extend to the brain and how they may attenuate cognitive dysfunction via modulating ROS after MI. After coronary ligation, male Wistar rats were randomized via an intramyocardial route to receive either vehicle, hADSC transplantation (1x106 cells), or the combination of hADSCs and 3-Morpholinosydnonimine (SIN-1, a peroxynitrite donor). Whether hADSCs migrated into the hippocampus was assessed by using human-specific primers in qPCR reactions. Passive avoidance test was used to assess cognitive performance. Postinfarction was associated with increased oxidative stress in myocardium, circulation and hippocampus. This was coupled with decreased numbers of dendritic spines as well as a significant downregulation of synaptic plasticity consisting of synaptophysin and PSD95. Step through latency during passive avoidance test was impaired in vehicle-treated rats after MI. Intramyocardial hADSC injection exerted therapeutic benefits in improving cardiac function and cognitive impairment. None of hADSCs were detected in rat’s hippocampus at the 3th day after intramyocardial injection. The beneficial effects of hADSCs on MI-induced histological and cognitive changes were abolished after adding SIN-1. MI-induced ROS attacked the hippocampus to induce neurodegeneration, resulting in cognitive deficit. The remotely intramyocardial administration of hADSCs, has the capacity of improved synaptic neuroplasticity in hippocampus mediated by ROS, not the cell engraftment, after MI.

scholarly journals The Use of Adipose-Derived Stem Cells (ADSCs) and Stromal Vascular Fraction (SVF) in Skin Scar Treatment—A Systematic Review of Clinical Studies

2021 ◽  
Vol 10 (16) ◽  
pp. 3637
Author(s):  
Albert Stachura ◽  
Wiktor Paskal ◽  
Weronika Pawlik ◽  
Maciej J. Mazurek ◽  
Janusz Jaworowski
Keyword(s):  
Systematic Review ◽  
Stem Cells ◽  
Platelet Rich Plasma ◽  
Stromal Vascular Fraction ◽  
Treatment Method ◽  
Adipose Derived Stem Cells ◽  
Beneficial Effects ◽  
Scar Treatment ◽  
Co2 Fractional Laser ◽  
Skin Scar

In recent years, lipofilling became a popular scar treatment method. Its beneficial outcomes have been partly attributed to the regenerative capacity of adipose-derived stem cells (ADSCs), suspended in an extracellular matrix—the stromal vascular fraction (SVF). The aim of this review was to verify if existing data support the clinical use of ADSC-related interventions in scar treatment. A systematic search of the literature was performed in July 2020 in five databases (Medline, Cochrane, Web of Science, Scopus and Embase). Articles written in English, except for reviews, letters and editorials, were identified and screened for eligibility. We looked for reports of any outcomes in scars treated with ADSCs or SVF. Data from selected articles were extracted and the quality of each study was assessed. Five hundred and fourteen studies were identified in the primary search, of which nineteen were eventually included in the systematic review. Extracted data pointed to beneficial microscopic, functional and aesthetic outcomes in a total of 665 patients. Six studies included comparative interventions—platelet-rich plasma or CO2 fractional laser. Collected data give low-to-average quality evidence for beneficial effects of ADSC-related interventions in scar treatment. Some studies suggest that these interventions are noninferior to PRP or fractional CO2 laser.

Adipose-Derived Stem Cells Improve Renal Function in a Mouse Model of IgA Nephropathy

2012 ◽  
Vol 21 (11) ◽  
pp. 2425-2439 ◽  
Author(s):  
Young Youl Hyun ◽  
In Ok Kim ◽  
Mi Hyung Kim ◽  
Deok Hwa Nam ◽  
Mi Hwa Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesangial Cells ◽  
Disease Onset ◽  
Immunoglobulin A ◽  
Adipose Derived Stem Cells ◽  
Beneficial Effects ◽  
Cytokine Balance ◽  
Before And After ◽  
Th1 Cytokine ◽  
Inflammatory Molecules

T-cell dysregulation plays an important role in the pathogenesis of immunoglobulin A nephropathy (IgAN). Adipose-derived stem cells (ASCs) have been reported to be able to prevent tissue damage through immune-modulating effects. To evaluate the effects of ASCs in high IgA ddY (HIGA) mice, ASCs were isolated from HIGA mice with different stages of IgAN before and after disease onset. ASCs were injected at a dose of 5 × 106 cells/kg body weight through the tail vein every 2 weeks for 3 months. Although the administered ASCs were rarely detected in the glomeruli, 24-h proteinuria was markedly decreased in all ASC-treated groups. Although glomerular deposition of IgA was not significantly different among groups, mesangial proliferation and glomerulosclerosis were dramatically decreased in most ASC treatment groups. In addition, levels of fibrotic and inflammatory molecules were markedly decreased by ASC treatment. Interestingly, ASC therapy significantly decreased Th1 cytokine activity in the kidney and caused a shift to Th2 responses in spleen T-cells as determined by FACS analysis. Furthermore, conditioned media from ASCs abrogated aggregated IgA-induced Th1 cytokine production in cultured HIGA mesangial cells. These results suggest that the beneficial effects of ASC treatment in IgAN occur via paracrine mechanisms that modulate the Th1/Th2 cytokine balance. ASCs are therefore a promising new therapeutic agent for the treatment of IgAN.

scholarly journals Technique for intraoperatory harvesting of adipose derived stem cells: towards cell treatment of recalcitrant ulcers

2017 ◽  
Vol 6 (2) ◽  
Author(s):  
Ilaria Zollino ◽  
Maria Grazia Sibilla ◽  
Sergio Gianesini ◽  
Erica Menegatti ◽  
Mirko Tessari ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Wound Healing ◽  
Adipose Tissue ◽  
Regenerative Medicine ◽  
Alternative Treatment ◽  
Treatment Options ◽  
Technical Note ◽  
Adipose Derived Stem Cells ◽  
Novel Therapy

Successful wound and ulcer repair remains a major biomedical challenge in the 21<sup>st</sup> Century. Innovative and alternative treatment options have been investigated over the last decade and stem cells application has been suggested as a possible novel therapy for regenerative medicine. In particular, stem cells derived from adipose tissue have been attracting a lot of attention in recent years as an alternative to the use of cells derived from bone marrow. This technical note describes the procedure introduced by Coleman for intraoperatory harvesting of adipose derived-stem-cells and proposes a rationale for using it in difficult wound healing and recalcitrant ulcers.

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