• Adipose-derived stem cells
• Dystrophinopathy
• Fibro-/adipogenic progenitors
• IL4
• Muscle fibrosis

• Mice
• Animals
• Interleukin-4
• Adipogenesis
• Cell Differentiation
• Stem Cells
• Satellite Cells, Skeletal Muscle
• Fibrosis
• Muscle, Skeletal/metabolism

• 81771359 Natural Science Foundation of China
• 81471280 Natural Science Foundation of China
• 2021A1515010313 Science and Technology Planning Project of Guangdong
• 2020B1212060017 Guangdong Provincial Key Laboratory of Diagnosis and Treatment of Major Neurological Diseases
• 2020B1111170002 Guangdong Provincial Clinical Research Center for Neurological Diseases of Chinese Medicine, Guangzhou University of Chinese Medicine
• 2015B050501003 Southern China International Joint Research Center for Early Intervention and Functional Rehabilitation of Neurological Diseases
• 2020A0505020004 Southern China International Joint Research Center for Early Intervention and Functional Rehabilitation of Neurological Diseases
• 201604020010 Guangdong Provincial Engineering Center for Major Neurological Disease Treatment, Guangdong Provincial Translational Medicine Innovation Platform for Diagnosis and Treatment of Major Neurological Disease, Guangzhou Clinical Research and Translational Cente

• Mechanobiology
• Skeletal muscle stem cells
• Stiffness
• Strain
• Topography
• Viscoelasticity

• C222812028 Biomedical Research Council
• OFYIRG21nov-0030 Agency for Science, Technology and Research
• A20D3b0073 Agency for Science, Technology and Research
• C210112057 Agency for Science, Technology and Research

The vocal cord in humans is essential in producing voice used in communication and interaction between us. Vocal cord paralysis causes dysphonia, which interferes with communication, causing disruptions towards social activity and daily activities. One of the managements for vocal cord paralysis is medialization and augmentation of the vocal cord through injection laryngoplasty. Autologous fat is one of the best fillers used in this procedure, but it is highly absorbable and can be reabsorbed very quickly when injected into body tissues. Platelet Rich Fibrin (PRF) is a biomaterial consisting of growth factors that are thought to improve fat tissue viability by increasing adipogenesis and angiogenesis. Improvement in fat viability will improve clinical outcomes after the laryngoplasty procedure, potentially reducing the number of repeated injections needed to achieve a satisfactory resolution to vocal cord paralysis. The study evaluates a combination of PRF and autologous microlobular fat compared with autologous microlobular fat alone on laryngoplasty. This single-blinded randomised control trial recruit a total of 18 patients, which are then randomised into the treatment and control groups. The evaluation was done via computerized acoustic analysis/Multidimensional Voice Program (MDVP) parameters and maximum phonation time. The MDVP results and maximum phonation time in both groups showed clinical improvement after the operation with no statistically significant differences.

•

PRF and PRP are platelet-derived products commonly used tissue engineering procedure.

• Autologous fat
• Injection laryngoplasty
• Platelet-rich fibrin
• Vocal cord paralysis

• RNA sequencing
• alkaline phosphatase
• gene expression
• human adipose-derived stromal/stem cells
• human dermal fibroblasts
• phenotypic characterization

Mesenchymal stromal/stem cells (MSCs) constitute a promising tool in regenerative medicine and can be isolated from different human tissues. However, their biological properties are still not fully characterized. Whereas MSCs from different tissue exhibit many common characteristics, their biological activity and some markers are different and depend on their tissue of origin. Understanding the factors that underlie MSC biology should constitute important points for consideration for researchers interested in clinical MSC application.

To characterize the biological activity of MSCs during longterm culture isolated from: bone marrow (BM-MSCs), adipose tissue (AT-MSCs), skeletal muscles (SM-MSCs), and skin (SK-MSCs).

MSCs were isolated from the tissues, cultured for 10 passages, and assessed for: phenotype with immunofluorescence and flow cytometry, multipotency with differentiation capacity for osteo-, chondro-, and adipogenesis, stemness markers with qPCR for mRNA for Sox2 and Oct4, and genetic stability for p53 and c-Myc; 27 bioactive factors were screened using the multiplex ELISA array, and spontaneous fusion involving a co-culture of SM-MSCs with BM-MSCs or AT-MSCs stained with PKH26 (red) or PKH67 (green) was performed.

All MSCs showed the basic MSC phenotype; however, their expression decreased during the follow-up period, as confirmed by fluorescence intensity. The examined MSCs express CD146 marker associated with proangiogenic properties; however their expression decreased in AT-MSCs and SM-MSCs, but was maintained in BM-MSCs. In contrast, in SK-MSCs CD146 expression increased in late passages. All MSCs, except BM-MSCs, expressed PW1, a marker associated with differentiation capacity and apoptosis. BM-MSCs and AT-MSCs expressed stemness markers Sox2 and Oct4 in long-term culture. All MSCs showed a stable p53 and c-Myc expression. BM-MSCs and AT-MSCs maintained their differentiation capacity during the follow-up period. In contrast, SK-MSCs and SM-MSCs had a limited ability to differentiate into adipocytes. BM-MSCs and AT-MSCs revealed similarities in phenotype maintenance, capacity for multilineage differentiation, and secretion of bioactive factors. Because AT-MSCs fused with SM-MSCs as effectively as BM-MSCs, AT-MSCs may constitute an alternative source for BM-MSCs.

Long-term culture affects the biological activity of MSCs obtained from various tissues. The source of MSCs and number of passages are important considerations in regenerative medicine.

• Mesenchymal stem/progenitor cells
• Bone marrow MSCs
• Adipose tissue MSCs
• Muscle-derived MSCs
• Skin-derived MSCs
• Cytokines and trophic factors of MSCs
• Spontaneous fusion of MSCs

• Blood analyses
• Duchenne Muscular Dystrophy
• Neutron Activation Analysis
• X-ray fluorescence
• hASCs therapy

The main purpose of this study is to evaluate, in vitro, the cytotoxicity of different commercial brands of hyaluronic acids to be used as a vehicle for injection of human adipose-derived mesenchymal stem cells (AD-MSCs).

AD-MSCs were divided into seven groups: one control group where AD-MSCs were cultivated with phosphate-buffered saline (PBS) and six other groups where AD-MSCs were cultivated with different commercial brands of hyaluronic acid. AD-MSC viability analysis was performed after 4, 24, and 48 h in contact with each treatment, using the trypan staining method on a Countess automated cell counter (Thermo Fisher Scientific).

The results clearly demonstrated a significant difference in cell viability when AD-MSCs were exposed to different hyaluronic acids when compared with the control group.

These data suggest that hyaluronic acid can be used as a vehicle for injection of human AD-MSCs, but caution is needed to choose the best product, aiming at its future therapeutic application.

• Arthroscopy
• Articular cartilage
• Cartilage diseases
• Knee
• Mesenchymal stem cell transplantation
• Mesenchymal stem cells

• Adipose-derived stem cells (ADSCs)
• Muscle regeneration therapy
• MyoD
• Myogenic differentiation

• Adipose-derived stem cells
• Muscle regeneration
• Myogenesis
• Porcine
• Skeletal muscle

• Adipose Tissue/cytology
• Animals
• Cell Differentiation
• Cell Lineage
• Cell Nucleus/metabolism
• Cell Separation
• Coculture Techniques
• Culture Media
• Gene Expression Regulation
• Mesenchymal Stem Cells/cytology
• Mice
• Muscle Development
• Muscle Fibers, Skeletal/metabolism
• Organ Specificity/genetics
• RNA, Messenger/genetics
• RNA, Messenger/metabolism
• Satellite Cells, Skeletal Muscle/metabolism
• Stem Cells/cytology
• Swine

• #2007-04072 Carle Foundation Hospital
• IDPH # 63080017 Illinois Regenerative Medicine Institute

Volumetric muscle loss (VML) injury resulted from massive muscle defects and diseases for which there are still no effective therapeutic treatments. This study aimed to investigate the effects of rat adipose-derived mesenchymal stem cells (rASCs) and rASCs-conditioned medium- (CM-) based type I collagen hydrogel on macrophage (MP) transition, myogenesis, and vascularization in the rat VML model. Laser Doppler results demonstrated much higher blood flow in the rASC- and CM-based hydrogel groups. qRT-PCR, hematoxylin and eosin, immunofluorescence, and Sirius Red staining manifested that both rASCs and CM-based hydrogel implantation accelerated muscle repair with upregulated angiogenesis and myogenesis, attenuated inflammation while facilitating M2 transition, and decreased the collagen deposition compared with the hydrogel group. In vitro experiments indicated that factors secreted from polarized M2 MPs could accelerate the migration and tube formation capacities of HUVECs. These results suggested that rASCs exerted immunomodulatory effects on MPs which further enhanced the proangiogenic potential on ECs to promote myogenesis and angiogenesis during muscle repair. These fundamental results support further clinical applications of ASCs for muscle loss injury.

• ADSC
• Reconstructive surgery
• Stem cells
• Tissue engineering

• Adipose Tissue/growth & development
• Animals
• Humans
• Plastic Surgery Procedures/methods
• Regeneration/physiology
• Stem Cell Transplantation/methods
• Stem Cells/physiology
• Surgery, Plastic/methods

Neurologists in their clinical practice are faced with inquiries about the suitability of stem cell approaches by patients with a variety of acute and chronic (namely neurodegenerative) disorders. The challenge is to provide these patients with accurate information about the scope of stem cell use as well as at the same time, empowering patients with the capacity to make an autonomous decision regarding the use of stem cells.

The Indian Academy of Neurology commissioned an Expert Group Meeting to formulate an advisory to practicing neurologists to counsel patients seeking information and advice about stem cell approaches.

In the course of such counselling, it should be emphasized that the information provided by many lay websites might be unsubstantiated. Besides, standard recommendations for the stem cell research, in particular, the application of several layers of oversight should be strictly adhered in order to ensure safety and ethical use of stem cells in neurological disorders.

• Cerebellar ataxias
• Parkinson's disease
• motor neuron disease
• multiple sclerosis
• muscle disorders
• neurological disorders
• spinal cord injury
• stem cells
• stroke

Adult skeletal muscle is a postmitotic tissue with an enormous capacity to regenerate upon injury. This is accomplished by resident stem cells, named satellite cells, which were identified more than 50 years ago. Since their discovery, many researchers have been concentrating efforts to answer questions about their origin and role in muscle development, the way they contribute to muscle regeneration, and their potential to cell-based therapies. Satellite cells are maintained in a quiescent state and upon requirement are activated, proliferating, and fusing with other cells to form or repair myofibers. In addition, they are able to self-renew and replenish the stem pool. Every phase of satellite cell activity is highly regulated and orchestrated by many molecules and signaling pathways; the elucidation of players and mechanisms involved in satellite cell biology is of extreme importance, being the first step to expose the crucial points that could be modulated to extract the optimal response from these cells in therapeutic strategies. Here, we review the basic aspects about satellite cells biology and briefly discuss recent findings about therapeutic attempts, trying to raise questions about how basic biology could provide a solid scaffold to more successful use of these cells in clinics.

Adipose-derived stem cells have been shown to promote peripheral nerve regeneration through the paracrine secretion of neurotrophic factors. However, it is unclear whether these cells can promote myogenic differentiation in muscular dystrophy. Adipose-derived stem cells (6 × 10⁶) were injected into the gastrocnemius muscle of mdx mice at various sites. Dystrophin expression was found in the muscle fibers. Phosphorylation levels of Akt, mammalian target of rapamycin (mTOR), eIF-4E binding protein 1 and S6 kinase 1 were increased, and the Akt/mTOR pathway was activated. Simultaneously, myogenin levels were increased, whereas cleaved caspase 3 and vimentin levels were decreased. Necrosis and fibrosis were reduced in the muscle fibers. These findings suggest that adipose-derived stem cells promote the regeneration and survival of muscle cells by inhibiting apoptosis and fibrosis, thereby alleviating muscle damage in muscular dystrophy.

• Duchenne muscular dystrophy
• adipose-derived stem cells
• dystrophin
• myogenic differentiation; paracrine pathway
• nerve regeneration
• neural regeneration

Pluripotent stem cells (PSCs), such as embryonic stem cells or induced pluripotent stem cells are a promising source of cells for regenerative medicine as they can differentiate into all cell types building a mammalian body. However, protocols leading to efficient and safe in vitro generation of desired cell types must be perfected before PSCs can be used in cell therapies or tissue engineering. In vivo, i.e. in developing mouse embryo or teratoma, PSCs can differentiate into skeletal muscle, but in vitro their spontaneous differentiation into myogenic cells is inefficient. Numerous attempts have been undertaken to enhance this process. Many of them involved mimicking the interactions occurring during embryonic myogenesis. The key regulators of embryonic myogenesis, such as Wnts proteins, fibroblast growth factor 2, and retinoic acid, have been tested to improve the frequency of in vitro myogenic differentiation of PSCs. This review summarizes the current state of the art, comparing spontaneous and directed myogenic differentiation of PSCs as well as the protocols developed this far to facilitate this process.

• Directed differentiation
• Myogenesis
• Pluripotent stem cells
• Regenerative medicine
• Skeletal muscle
• Spontaneous differentiation

• Animals
• Cell Differentiation/physiology
• Humans
• Muscle Development/physiology
• Muscle, Skeletal/physiology
• Pluripotent Stem Cells/physiology

• Fundacja na rzecz Nauki Polskiej (PL)
• Narodowe Centrum Nauki (PL)

Since the discovery of adipose-derived stem cells (ASC) in human adipose tissue nearly 15 years ago, significant advances have been made in progressing this promising cell therapy tool from the laboratory bench to bedside usage. Standardization of nomenclature around the different cell types used is finally being adopted, which facilitates comparison of results between research groups. In vitro studies have assessed the ability of ASC to undergo mesenchymal differentiation as well as differentiation along alternate lineages (transdifferentiation). Recently, focus has shifted to the immune modulatory and paracrine effects of transplanted ASC, with growing interest in the ASC secretome as a source of clinical effect. Bedside use of ASC is advancing alongside basic research. An increasing number of safety-focused Phase I and Phase IIb trials have been published without identifying any significant risks or adverse events in the short term. Phase III trials to assess efficacy are currently underway. In many countries, regulatory frameworks are being developed to monitor their use and assure their safety. As many trials rely on ASC injected at a distant site from the area of clinical need, strategies to improve the homing and efficacy of transplanted cells are also being explored. This review highlights each of these aspects of the bench-to-bedside use of ASC and summarizes their clinical utility across a variety of medical specialties.

• bystander effect
• mesenchymal stem cells
• standardization
• stromal cells
• stromal vascular fraction

Regenerative capacity of skeletal muscles resides in satellite cells, a self-renewing population of muscle cells. Several studies are investigating epigenetic mechanisms that control myogenic proliferation and differentiation to find new approaches that could boost regeneration of endogenous myogenic progenitor populations. In recent years, a lot of effort has been applied to purify, expand and manipulate adult stem cells from muscle tissue. However, this population of endogenous myogenic progenitors in adults is limited and their access is difficult and invasive. Therefore, other sources of stem cells with potential to regenerate muscles need to be examined. An excellent candidate could be a population of adult stromal cells within fat characterized by mesenchymal properties, which have been termed adipose-derived stem cells (ASCs). These progenitor adult stem cells have been successfully differentiated in vitro to osteogenic, chondrogenic, neurogenic and myogenic lineages. Autologous ASCs are multipotent and can be harvested with low morbidity; thus, they hold promise for a range of therapeutic applications. This review will summarize the use of ASCs in muscle regenerative approaches.

• adipose
• muscle
• regeneration
• reprogramming
• stem cells
• transdifferentiation
• transplantation

The use of Mesenchymal Stromal Cells (MSCs) aiming to treat cancer has shown very contradictory results. In an attempt to clarify the contradictory results reported in the literature and the possible role of human fallopian tube Mesenchymal Stromal Cells (htMSCs) against breast cancer, the aim of this study was to evaluate the clinical effect of htMSCs in murine mammary adenocarcinoma using two different approaches: (1) coinjections of htMSCs and 4T1 murine tumor cell lineage and (2) injections of htMSCs in mice at the initial stage of mammary adenocarcinoma development. Coinjected animals had a more severe course of the disease and a reduced survival, while tumor-bearing animals treated with 2 intraperitoneal injections of 10⁶ htMSCs showed significantly reduced tumor growth and increased lifespan as compared with control animals. Coculture of htMSCs and 4T1 tumor cells revealed an increase in IL-8 and MCP-1 and decreased VEGF production. For the first time, we show that MSCs isolated from a single source and donor when injected in the same animal model and tumor can lead to opposite results depending on the experimental protocol. Also, our results demonstrated that htMSCs can have an inhibitory effect on the development of murine mammary adenocarcinoma.

• Adipose derived mesenchymal stem cells,
• Anxiety
• Huntington’s disease,
• Locomotor activity,

• Adipose Tissue/cytology
• Animals
• Anxiety
• Corpus Striatum/metabolism
• Corpus Striatum/pathology
• Corpus Striatum/surgery
• Disease Models, Animal
• Humans
• Huntington Disease/pathology
• Huntington Disease/psychology
• Huntington Disease/surgery
• Male
• Mesenchymal Stem Cell Transplantation/methods
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Motor Activity
• Rats
• Rats, Wistar
• Rotarod Performance Test
• Transplantation, Heterologous/methods

• Adult stem cell therapy
• Cellular therapy
• Mesenchymal stromal cells
• Stem cell research
• Stress urinary incontinence

Human amniotic fluid stem (hAFS) cells have been shown to differentiate into multiple lineages, including myoblasts. However, molecular mechanisms underlying the myogenic differentiation of hAFS cells and their regenerative potential for muscle injury remain to be elucidated.

In order to induce myogenic differentiation of hAFS cells, lentiviruses for MYOD were constructed and transduced into hAFS cells. Formation of myotube-like cells was analyzed by immunocytochemistry, and expression of molecular markers for myoblasts was analyzed by reverse transcription polymerase chain reaction and Western blotting. For in vivo muscle regeneration, MYOD transduced hAFS cells were injected into left tibialis anterior (TA) muscles injured with cardiotoxin, and muscle regeneration was analyzed using hematoxylin and eosin, immunocytochemistry and formation of neuro-muscular junction.

MYOD expression in hAFS cells successfully induced differentiation into multinucleated myotube-like cells. Consistently, significant expression of myogenic marker genes, such as MYOG, DES, DMD and MYH, was induced by MYOD. Analysis of pre-myogenic factors showed that expression of PAX3, MEOX1 and EYA2 was significantly increased by MYOD. MYOD was phosphorylated and localized in the nucleus. These results suggest that in hAFS cells, MYOD is phosphorylated and localized in the nucleus, thus inducing expression of myogenic factors, resulting in myogenic differentiation of hAFS cells. To test regenerative potential of MYOD-transduced hAFS cells, we transplanted them into injured muscles of immunodeficient BALB/cSlc-nu mice. The results showed a substantial increase in the volume of TA muscle injected with MYOD-hAFS cells. In addition, TA muscle tissue injected with MYOD-hAFS cells has more numbers of neuro-muscular junctions compared to controls, indicating functional restoration of muscle injury by MYOD-hAFS cells.

Collectively, our data suggest that transduction of hAFS cells with MYOD lentiviruses induces skeletal myogenic differentiation in vitro and morphological and functional regeneration of injured muscle in vivo.

Myostatin, a TGF-β family member, is associated with inhibition of muscle growth and differentiation and might interact with the IGF-1 signaling pathway. Since IGF-1 is secreted at a bioactive level by adipose tissue-derived mesenchymal stem cells (ASCs), these cells (ASCs) provide a therapeutic option for Duchenne Muscular Dystrophy (DMD). But the protective effect of stem cell secreted IGF-1 on myoblast under high level of myostatin remains unclear. In the present study murine myoblasts were exposed to myostatin under presence of ASCs conditioned medium and investigated for proliferation and apoptosis. The protective effect of IGF-1 was further examined by using IGF-1 neutralizing and receptor antibodies as well as gene silencing RNAi technology. MyoD expression was detected to identify impact of IGF-1 on myoblasts differentiation when exposed to myostatin. IGF-1 was accountable for 43.6% of the antiapoptotic impact and 48.8% for the proliferative effect of ASCs conditioned medium. Furthermore, IGF-1 restored mRNA and protein MyoD expression of myoblasts under risk. Beside fusion and transdifferentiation the beneficial effect of ASCs is mediated by paracrine secreted cytokines, particularly IGF-1. The present study underlines the potential of ASCs as a therapeutic option for Duchenne muscular dystrophy and other dystrophic muscle diseases.

• Adipose Tissue/cytology
• Adult
• Cell Communication
• Cell Culture Techniques/instrumentation
• Cell Differentiation
• Cell Membrane/metabolism
• Cell Movement
• Cell Proliferation
• Coculture Techniques/instrumentation
• Elastic Modulus
• Equipment Design
• Extracellular Matrix/metabolism
• Gels/chemistry
• Humans
• Muscle Development
• Myoblasts/cytology
• Pressure
• Silicon/chemistry
• Stem Cells/cytology
• Young Adult

• T32 EB009380 NIBIB NIH HHS
• 1-DP2-OD004309 NIH HHS
• DP2 OD006460 NIH HHS
• T32 HL105373 NHLBI NIH HHS
• T32 HD060555 NICHD NIH HHS
• 1-DP2-OD006460 NIH HHS
• R01 GM132106 NIGMS NIH HHS
• DP2 OD004309 NIH HHS

• Adipocytes/metabolism
• Animals
• Cell Differentiation/drug effects
• Cell Differentiation/physiology
• Culture Techniques
• Epithelial Cells/cytology
• Epithelial Cells/metabolism
• Female
• Fibroblast Growth Factor 7/pharmacology
• Gene Expression Regulation/drug effects
• Goats/growth & development
• Goats/metabolism
• Lipoprotein Lipase/metabolism
• Mammary Glands, Animal/cytology
• Mammary Glands, Animal/metabolism
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/drug effects
• PPAR gamma/metabolism
• Resistin/metabolism
• Sexual Maturation
• Thiazolidinediones/pharmacology
• alpha-Linolenic Acid/pharmacology

• Adipose Tissue/metabolism
• Adipose Tissue/pathology
• Biocompatible Materials/chemistry
• Cell Adhesion
• Cell Differentiation
• Cell Proliferation
• Humans
• Immunohistochemistry
• Microscopy, Electron, Scanning
• Muscle Development
• Muscle, Skeletal/metabolism
• Myosins/metabolism
• Nanofibers/chemistry
• Nanostructures/chemistry
• Nanotechnology
• Polyesters/chemistry
• Stem Cells/cytology
• Tissue Engineering/methods
• Tissue Scaffolds/chemistry
• Tropomyosin/chemistry

The stromal-vascular cell fraction (SVF) of adipose tissue can be an abundant source of both multipotent and pluripotent stem cells, known as adipose-derived stem cells or adipose tissue-derived stromal cells (ADSCs). The SVF also contains vascular cells, targeted progenitor cells, and preadipocytes. Stromal cells isolated from adipose tissue express common surface antigens, show the ability to adhere to plastic, and produce forms that resemble fibroblasts. They are characterized by a high proliferation potential and the ability to differentiate into cells of meso-, ecto- and endodermal origin. Although stem cells obtained from an adult organism have smaller capabilities for differentiation in comparison to embryonic and induced pluripotent stem cells (iPSs), the cost of obtaining them is significantly lower. The 40 years of research that mainly focused on the potential of bone marrow stem cells (BMSCs) revealed a number of negative factors: the painful sampling procedure, frequent complications, and small cell yield. The number of stem cells in adipose tissue is relatively large, and obtaining them is less invasive. Sampling through simple procedures such as liposuction performed under local anesthesia is less painful, ensuring patient comfort. The isolated cells are easily grown in culture, and they retain their properties over many passages. That is why adipose tissue has recently been treated as an attractive alternative source of stem cells. Essential aspects of ADSC biology and their use in regenerative medicine will be analyzed in this article.

• cell migration
• cell survival
• immune privilege
• mesenchymal stromal cells
• stem cell therapy
• xenotransplantation

• Xenotransplantation
• blood-retina barrier
• cell integration
• cell migration
• cell survival
• stem-cell therapy

Osteoarticular pathologies very often require an implementation therapy to favor regeneration processes of bone, cartilage and/or tendons. Clinical approaches performed on osteoarticular complications in dogs constitute an ideal model for human clinical translational applications. The adipose-derived mesenchymal stem cells (ASCs) have already been used to accelerate and facilitate the regenerative process. ASCs can be maintained in vitro and they can be differentiated to osteocytes or chondrocytes offering a good tool for cell replacement therapies in human and veterinary medicine. Although ACSs can be easily obtained from adipose tissue, the amplification process is usually performed by a time consuming process of successive passages. In this work, we use canine ASCs obtained by using a Bioreactor device under GMP cell culture conditions that produces a minimum of 30 million cells within 2 weeks. This method provides a rapid and aseptic method for production of sufficient stem cells with potential further use in clinical applications. We show that plasma rich in growth factors (PRGF) treatment positively contributes to viability and proliferation of canine ASCs into caprolactone 2-(methacryloyloxy) ethyl ester (CLMA) scaffolds. This biomaterial does not need additional modifications for cASCs attachment and proliferation. Here we propose a framework based on a combination of approaches that may contribute to increase the therapeutical capability of stem cells by the use of PRGF and compatible biomaterials for bone and connective tissue regeneration.

• Actin Cytoskeleton/physiology
• Bone Marrow Cells/cytology
• Cell Differentiation/drug effects
• Humans
• Insulin-Like Growth Factor I/pharmacology
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Muscle, Skeletal/cytology
• Muscle, Skeletal/metabolism
• MyoD Protein/genetics
• MyoD Protein/metabolism
• Myogenic Regulatory Factor 5/genetics
• Myogenic Regulatory Factor 5/metabolism
• Myogenic Regulatory Factors/genetics
• Myogenic Regulatory Factors/metabolism
• Myogenin/genetics
• Myogenin/metabolism
• Stress, Mechanical

• fallopian tube
• mesenchymal stromal cells
• bone regeneration
• regenerative medicine
• xenotransplant

• Adult
• Animals
• Bone Regeneration/physiology
• Cell Differentiation
• Cell Lineage
• DNA/metabolism
• Fallopian Tubes/cytology
• Female
• Flow Cytometry
• Humans
• Male
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/ultrastructure
• Middle Aged
• Models, Biological
• Osteogenesis
• Rats
• Rats, Wistar
• Transplantation, Heterologous