Role Of Cell Therapy Research Articles

High-density culturing of the dermal fibroblast cells on hydrogel-based soft microcarriers for cell therapy application

Dermal fibroblasts play a pivotal role in cell therapy, serving as versatile cells essential for tissue repair and regeneration. Their unique ability to produce extracellular matrix components contributes significantly to the efficacy and success of various cell-based therapies in treating skin-related conditions and wounds. Scaling up the production of dermal fibroblasts poses a challenge due to their limited lifespan in culture and the need to maintain their specialized functions over multiple generations. Bioreactors offer a promising solution by providing controlled environments that optimize cell growth, allowing for higher yields of dermal fibroblasts while preserving their functionality and minimizing the limitations associated with traditional culture methods. To overcome the critical challenges in large-scale cell manufacturing, we fine-tuned the composition of our existing smart microcarrier, Cytogel, and tailored its chemical and physical properties to cultivate dermal fibroblasts. The fibroblast cell attachment rate to the new optimized formulation is more than 90%, followed by 60% thermal detachment efficiency in 1 h. Additionally, cells harvested from Cytogel displayed superior CD90 expression and enhanced collagen production compared to fibroblasts cultured in tissue flasks in both static and dynamic bioreactor cultures. Overall, our developed hydrogel-based microcarriers exhibited significant promise for efficient cell expansion, offering a groundbreaking approach for industrial-scale cell culture for cell therapeutic applications.

Proteomic analysis and functional validation reveal distinct therapeutic capabilities related to priming of mesenchymal stromal/stem cells with IFN-γ and hypoxia: potential implications for their clinical use.

Mesenchymal stromal/stem cells (MSCs) are a heterogeneous population of multipotent cells that can be obtained from various tissues, such as dental pulp, adipose tissue, bone marrow and placenta. MSCs have gained importance in the field of regenerative medicine because of their promising role in cell therapy and their regulatory abilities in tissue repair and regeneration. However, a better characterization of these cells and their products is necessary to further potentiate their clinical application. In this study, we used unbiased high-resolution mass spectrometry-based proteomic analysis to investigate the impact of distinct priming strategies, such as hypoxia and IFN-γ treatment, on the composition and therapeutic functionality of the secretome produced by MSCs derived from the amniotic membrane of the human placenta (hAMSCs). Our investigation revealed that both types of priming improved the therapeutic efficacy of hAMSCs, and these improvements were related to the secretion of functional factors present in the conditioned medium (CM) and exosomes (EXOs), which play crucial roles in mediating the paracrine effects of MSCs. In particular, hypoxia was able to induce a pro-angiogenic, innate immune response-activating, and tissue-regenerative hAMSC phenotype, as highlighted by the elevated production of regulatory factors such as VEGFA, PDGFRB, ANGPTL4, ENG, GRO-γ, IL8, and GRO-α. IFN-γ priming, instead, led to an immunosuppressive profile in hAMSCs, as indicated by increased levels of TGFB1, ANXA1, THBS1, HOMER2, GRN, TOLLIP and MCP-1. Functional assays validated the increased angiogenic properties of hypoxic hAMSCs and the enhanced immunosuppressive activity of IFN-γ-treated hAMSCs. This study extends beyond the direct priming effects on hAMSCs, demonstrating that hypoxia and IFN-γ can influence the functional characteristics of hAMSC-derived secretomes, which, in turn, orchestrate the production of functional factors by peripheral blood cells. This research provides valuable insights into the optimization of MSC-based therapies by systematically assessing and comparing the priming type-specific functional features of hAMSCs. These findings highlight new strategies for enhancing the therapeutic efficacy of MSCs, particularly in the context of multifactorial diseases, paving the way for the use of hAMSC-derived products in clinical practice.

MICROSCOPIC CHANGES IN MUSCLES OF RABBITS WITH EXPERIMENTALLY DAMAGED MUSCLE TISSUE UNDER THE INFLUENCE OF TRANSPLANTED MESENCHYMAL STEM CELLS

Due to the high level of activity of some animals, particularly sports and working animals, muscle injuries are becoming a serious and urgent problem for veterinarians. Sporting events, training and work duties can lead to tears, strains and other injuries to skeletal muscle tissue. Muscle injuries in sports and working animals are a complex problem that requires a careful and comprehensive approach to treatment. These injuries can lead to pain, restricted movement and loss of performance. Traditional treatments, while effective, are often lengthy and not always successful. Therefore, scientists and veterinarians are constantly looking for new and more effective ways to treat and rehabilitate animals to help them recover from injuries faster and more fully. The main purpose of this article is to review, summarise and analyse histological studies investigating the use of allogeneic mesenchymal stem cells in veterinary medicine for regenerative therapy. An additional goal of the article is to review the results of these studies and highlight the main conclusions regarding the potential use of MSCs in veterinary practice. The main role in cell therapy is played by allogeneic mesenchymal stem cells (MSCs). These cells, which do not have a specific differentiation, can independently renew themselves and develop into cells with specific functions, making them effective for tissue repair. It is especially important to note that mesenchymal stem cells are promising for veterinary medicine because they are easy to isolate and cultivate. The article discusses the use of allogeneic mesenchymal stem cells for tissue repair in veterinary medicine in experimental animals, taking into account the results of scientific research conducted by the authors of the article. The use of a new and promising method of treating animals with damaged muscle tissue based on cellular regeneration can significantly accelerate the recovery of animals after injury and avoid side effects that often occur with traditional treatments.

Acetyl l-carnitine protects adipose-derived stem cells against serum-starvation: regulation on the network composed of reactive oxygen species, autophagy, apoptosis and senescence.

Adipose-derived stem cells (ADSCs) play an important role in cell therapy and regenerative medicine. However, local nutritional deficiency often limits therapeutical effect of the transplanted cells. Acetyl l-carnitine (ALC) is a common energy metabolism regulator and free radical scavenger. This study investigated the effect of ALC on ADSCs exposed to severe serum-deprivation and explored the relative machanisms. Treating with 1mM ALC improved proliferation and alleviated senescence of starved cells, accompanied with reduced reactive oxygen species (ROS) and increased protein expression of SOD1 and catalase. In addition, ALC inhibited apoptosis but increased starvation-induced autophagy, which might be related to the regulation of phases of dissociation of Bcl-2-Beclin1 and Bcl-2-Bax complexes. Evidence obtained by replacing ALC with N-acetylcysteine (N-AC) suggested that ROS might be the central inducer of autophagy, apoptosis and senescence. There was a difference between ALC and N-AC in the protection mechanism, that was, compared with N-AC, ALC maintained autophagy well at the same time as anti-oxidation. Inhibition of autophagy by 3-methyladenine (3-MA) partially offset the protective effect of ALC. However, despite low-level ROS and enhanced autophagy, ALC with high concentration (10mM) markedly aggravated cell apoptosis and senescence, thus losing cytoprotection and even causing damage.

Neural stem cells as a promising therapeutic approach in neurological diseases

The central nervous system lacks the ability to undergo neuronal regeneration. Neurological diseases such as neurotrauma and neurovascular diseases impact many people around the world. Neural stem cell-based treatment for neurological diseases limits clinical damage by providing and by regeneration local support for damaged and lost neural cells. Neural stem cells (NSCs) have a vital role in cell therapy as they can give rise to neurons or glial cells and repair neurological injuries. NSCs were able to secrete constitutively neuroprotective factors that have an important role in neurological repair. Moreover, upon transplantation, NSCs were shown capable of migration into lesioned sites and improved functional recovery after neurological diseases due to their important role in neurogenesis, immunomodulation, and brain plasticity. This review focuses on the roles of NSCs-based therapies in neurological diseases and summarise current advancements in NSC transplantation into rodent models of neurological disorders as well as shed the light on the progress of NSCs therapies in clinical trials.

Potential Mechanisms and Perspectives in Ischemic Stroke Treatment Using Stem Cell Therapies

Ischemic stroke (IS) remains one of the major causes of death and disability due to the limited ability of central nervous system cells to regenerate and differentiate. Although several advances have been made in stroke therapies in the last decades, there are only a few approaches available to improve IS outcome. In the acute phase of IS, mechanical thrombectomy and the administration of tissue plasminogen activator have been widely used, while aspirin or clopidogrel represents the main therapy used in the subacute or chronic phase. However, in most cases, stroke patients fail to achieve satisfactory functional recovery under the treatments mentioned above. Recently, cell therapy, especially stem cell therapy, has been considered as a novel and potential therapeutic strategy to improve stroke outcome through mechanisms, including cell differentiation, cell replacement, immunomodulation, neural circuit reconstruction, and protective factor release. Different stem cell types, such as mesenchymal stem cells, marrow mononuclear cells, and neural stem cells, have also been considered for stroke therapy. In recent years, many clinical and preclinical studies on cell therapy have been carried out, and numerous results have shown that cell therapy has bright prospects in the treatment of stroke. However, some cell therapy issues are not yet fully understood, such as its optimal parameters including cell type choice, cell doses, and injection routes; therefore, a closer relationship between basic and clinical research is needed. In this review, the role of cell therapy in stroke treatment and its mechanisms was summarized, as well as the function of different stem cell types in stroke treatment and the clinical trials using stem cell therapy to cure stroke, to reveal future insights on stroke-related cell therapy, and to guide further studies.

Astaxanthin protects mesenchymal stem cells from oxidative stress by direct scavenging of free radicals and modulation of cell signaling

Recent evidence has shown that mesenchymal stem cells (MSCs) play vital roles in cell therapy of ischemia/hypoxia damaged tissues. However, after the transplantation, they might undergo apoptosis due to oxidative stress. Thus, some strategies have been developed to support stem cells in harsh conditions, including pre-treatment of the cells with antioxidants. Of various antioxidants, in this study, astaxanthin (ATX) was used to protect adipose-derived MSCs against oxidative stress. The MSCs were exposed to different doses of hydrogen peroxide, and then the expression of key genes involved in the redox signaling pathway was studied, including nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADPH quinine oxidoreductase 1 (NQO1). The balance of intracellular reactive oxygen species was detected with the H2DCFDA molecular probe. Additionally, for the detection of apoptosis and protective effect of ATX, the DAPI/Phallacidin and annexin V cell staining were performed. The results of cellular studies revealed that ATX reduced the H2O2-induced cell apoptosis and oxidative stress. Furthermore, after the induction of oxidative stress, the cells’ native antioxidants (HO-1 and NQO1) were overexpressed but they were modulated with ATX treatments (p < 0.023). Based on our findings, ATX could increase the expression of Nrf2 as a key transcription factor of antioxidant enzymes (p < 0.05). These findings support the notion that ATX can act as an effective antioxidant in the pre-treatment of MSCs before cell therapy. Thus, to enhance the viability of stem cells during the transplantation in harsh conditions, the concurrent use of ATX in cell therapy modalities is proposed.

Isolation and biological characteristics of sheep amniotic epithelial cells.

Amniotic epithelial cells (AECs), isolated from placenta, have epithelial cells and stem cells characteristics. Most of the previous studies focused on the biological characteristics of human amniotic epithelial cells, which demonstrated amniotic epithelial cells not only had low immunogenicity and potent potential to differentiate into three germ layers, but also could secrete various immunomodulatory factors. However, compared to study on human amniotic epithelial cells, few studies have been done on other animals. In this study, sheep amniotic epithelial cells were successfully isolated and their surface makers were accessed by immunofluorescence assay, and found that AECs were expressed Oct4 and Sox2, which were necessary for maintaining the undifferentiated state of pluripotent stem cells. Based on cloning efficiency and growth kinetics assay, AECs were found to possess self-renewal capacity and the growth curve was S-shaped. In addition, AECs could be induced into adipocytes, osteoblasts and chondrocytes in vitro, showing they had multi-differential ability. Reverse transcription-polymerase chain reaction results showed that AECs expressed CD29, CD44, CD90 and CK19, and didn't expressed CD34, CD45 and the telomerase gene (TERT). Little change in chromosome number was observed in AEC cultures for up to at least the first ten passages. In summary, this study results revealed that sheep AECs possessed more advantages for cell therapy and might play a key role in cell therapy and regenerative medicine in the future.

Cumulus Cells Are Potential Candidates for Cell Therapy.

Cumulus cells (CCs) originate from the membrane granulosa cells and surround oocytes during follicle maturation. CCs produce high levels of hyaluronan that targets CD44, which is a major tumorigenic marker. This study aimed to investigate whether CCs have a role in cell therapy by targeting CD44 in pancreatic cancer cells. CCs were isolated from the oocytes and incubated in a hypoxic environment. BxPC-3 pancreatic cancer cells were treated with CC conditioned media for three days. Conditioned media of CC cells incubated in hypoxic conditions caused a 25% reduction in the viability of BxPC-3 cells. Expression of anti-apoptotic genes was down-regulated, while that of pro-apoptotic genes was upregulated. An increased number of BxPC-3 cells exhibited increased levels of reactive oxygen species and arrested in the synthesis (S) phase of the cell cycle. CCs conditioned medium induced apoptosis of pancreatic cancer cells.

The Role of Cell Therapy in Treating Cancer

Abstract: With the continuous development and advancement in the field of medical technology, cell therapy for treating cancer has received extensive attention. Compared with conventional therapy, cell therapy provides a new direction of scientific research, which is worthy for clinical application and promotion. This article focuses on the role of cell therapy in cancer treatment and possibly a good reference for the future.

The role of cell therapies and hip arthroscopy in the management of osteonecrosis: an update.

ABSTRACTThe exact pathophysiology of osteonecrosis of the femoral head (ONFH) is still unknown. There is evidence to suggest that in ON there is decreased population and altered function of the mesenchymal stem cells (MSCs) of the femoral head. This could influence both the actual occurrence of ON itself and the repair process that follows. Hence, in such an environment it only is rational to consider the use of cell-based treatments to potentially regenerate lost or damaged bone. The aim of this review is to provide an up-to-date, evidence-based information in the use of cell therapies in the treatment of nontraumatic ONFH and the use of hip arthroscopy in the field.

Therapeutic strategies involving uterine stem cells in reproductive medicine.

The current review provides an update on recent advances in stem cell biology relevant to female reproduction. Stem cells are undifferentiated cells that often serve as a reservoir of cells to regenerate tissue in settings or injury or cell loss. The endometrium has progenitor stem cells that can replace all of the endometrium during each menstrual cycle. In addition, multipotent endometrial cells replace these progenitor cells when depleted. Recruitment of stem cells from outside of the uterus occurs in setting of increased demand such as ischemia or injury. Bone marrow-derived multipotent stem cells are recruited to the uterus by estrogen or injury-induced expression of the chemokine CXCL12. In the setting of overwhelming injury, especially in the setting of low estrogen levels, there may be insufficient stem cell recruitment to adequately repair the uterus resulting in conditions such as Asherman syndrome or other endometrial defects. In contrast, excessive recruitment of stem cells underlies endometriosis. Enhanced understanding of stem-cell mobilization, recruitment, and engraftment has created the possibility of improved therapy for endometrial defects and endometriosis through enhanced manipulation of stem-cell trafficking. Further, the normal endometrium is a rich source of multipotent stem cells that can be used for numerous applications in regenerative medicine beyond reproduction. A better understanding of reproductive stem-cell biology may allow improved treatment of endometrial disease such as Asherman syndrome and other endometrial receptivity defects. Inhibiting stem-cell mobilization may also be helpful in endometriosis therapy. Finally, endometrial derived multipotent stem cells may play a crucial role in cell therapy for regenerative medicine.

Trehalose Activates Autophagy and Prevents Hydrogen Peroxide-Induced Apoptosis in the Bone Marrow Stromal Cells.

Bone marrow stromal stem cells (BMSCs) play a significant role in cell therapy. These cells quickly die after transplantation to the affected area due to oxidative stress. The natural disaccharide, trehalose which can be known as autophagy inducer. The present study aimed to investigate the role of trehalose in preventing BMSCs from oxidative stress caused by H2O2. BMSCs were isolated from the adult rats. The cells were divided into three groups: (a) control; (b) 100 µM H2O2; (c) 100 µM H2O2 and trehalose 3%. The morality rate was analyzed by viability test. Immunocytochemistry and Western blot was used in order to evaluate p62 protein and LC3II/LC3I ratio, respectively. In order to evaluate apoptosis, cleaved caspase-3 protein was used. In viability test, the survival rate for BMSCs after 8 h were 82%, 72%, 49%, and 39% (for groups who received 50, 100, 200, and 400 µM H2O2, respectively) compared to the control group. Pre-treatment with the use of trehalose 3% increased cell survivals. The levels of p62 protein, were increased in the cells under H2O2 treatment, while the levels of p62 protein in the cytoplasm, as autophagy inclusions, reduced for the group with trehalose pre-treatment. In addition, trehalose caused to increase LC3II/LC3I ratio and decreased the expression of cleaved caspase-3. Trehalose decreased apoptosis and increased the autophagy and survival levels of the cells against H2O2. Due to the unique properties of trehalose and its low toxicity, it can be used as a pharmaceutical agent in cellular transplantation to reduce oxidative stress.

Cell-Based Therapy in Alzheimer's Disease: Current Knowledge and Perspective

Alzheimer's disease (AD) is the most prevalent type of dementia, and its neuropathology is characterized by the deposition of insoluble β-amyloid (Aβ) peptides, intracellular neurofibrillary tangles, amyloid angiopathy, age-related brain atrophy, synaptic pathology, white matter rarefaction, granulovacuolar degeneration, neuron loss, and neuroinflammation. Although much is known about the neurobiology of AD, very few conventional therapies are available to arrest or slow the disease. There is an urgent need for novel therapeutic approaches for AD. AD subjects have significantly fewer viable precursor cells in the hippocampus compared with age-matched healthy control subjects. However, the viable precursor cells that remain in AD and age-matched healthy control brain specimens can be induced to differentiate. To facilitate or mimic the natural compensatory effect in AD, cell therapy, including endogenous and exogenous stem cells, has been considered in AD. In this review, we focus on the history and development of cell therapy in AD, and consider the role of cell therapy as a potential treatment for AD.

Application of Allogeneic Fibroblast Cells in Cellular Therapy of Recessive Dystrophic Epidermolysis Bullosa

Context: Connective tissue cells include fibroblasts, chondrocytes, adipocyte, and osteocytes. These cells are specialized for the secretion of collagenous extracellular matrix and are responsible for the architectural framework of the human body. Evidence Acquisition: Connective tissue cells play a central role in supporting as well as repairing tissues and organs. Fibroblast cell therapy could be used for the treatment of burn wounds, scars, diabetic foot ulcers, acne scars and skin aging. This review focused on biology of fibroblasts and their role in cell therapy of recessive dystrophic epidermolysis bullosa (RDEB). Results: Fibroblasts are known to play a pivotal role in skin structure and integrity, and dermal fibroblasts are believed to promote skin regeneration and rejuvenation via collagen production. Conclusions: Fibroblasts can be used in transplantations to ameliorate an immune system response, in order to reduce antigen production. Human fibroblasts suppress ongoing mixed lymphocyte reactions (MLRs) between lymphocyte cells from two individuals, and supernatant materials from fibroblast cultures suppress MLRs.

Automation Highlights from the Literature

Automation Highlights from the Literature

Cell selective chitosan microparticles as injectable cell carriers for tissue regeneration

The detection, isolation and sorting of cells holds an important role in cell therapy and regenerative medicine. Also, injectable systems have been explored for tissue regeneration in vivo, because it allows repairing complex shaped tissue defects through minimally invasive surgical procedures. Here we report the development of chitosan microparticles with a size of 115.8 μm able to capture and expand a specific cell type that can also be regarded as an injectable biomaterial. Monoclonal antibodies against cell surface antigens specific to endothelial cells and stem cells were immobilized on the surface of the microparticles. Experimental results showed that particles bioconjugated with specific antibodies provide suitable surfaces to capture a target cell type and subsequent expansion of the captured cells. Primarily designed for an application in tissue engineering, three main challenges are accomplished with the herein presented microparticles: separation, scale-up expansion of specific cell type and successful use as an injectable system to form small tissue constructs in situ.

Platelet lysates and their role in cell therapy

There is an emerging field of application for human platelet concentrates as they are rich in growth factors for in vitro culture of various cell types. To this end, the platelets are lysed to release the growth factors to the culture medium, for example by freeze–thaw procedures or by ultrasonic treatment. This human platelet lysate (HPL) is advantageous as compared to the traditionally used foetal bovine serum (FBS): it is xenogen‐free – and hence suitable for generation of therapeutic products – and it further increases proliferation rates of cells in culture. On the other hand, neither HPL nor FBS are precisely defined, and there is notorious batch‐to‐batch variation. Further research is required to unravel the relevant compounds to generate fully defined synthetic culture media – until then, HPL provides a suitable and reliable cell culture supplement particularly in cellular therapy.

CELL THERAPY IN THE TREATMENT OF CRITICAL LOWER LIMB ISCHEMIA

In recent years, the role of cell therapy in the treatment of patients with atherosclerotic severe lower limb ischemia is increased. The aim of our study was to compare the efficacy of non-viral fibroblast grow factor (NV1FGF) and autologous adipose stem cells (ASC) derived from subcutaneous fat in

Large-scale cell production of stem cells for clinical application using the automated cell processing machine

BackgroundCell-based regeneration therapies have great potential for application in new areas in clinical medicine, although some obstacles still remain to be overcome for a wide range of clinical applications. One major impediment is the difficulty in large-scale production of cells of interest with reproducibility. Current protocols of cell therapy require a time-consuming and laborious manual process. To solve this problem, we focused on the robotics of an automated and high-throughput cell culture system. Automated robotic cultivation of stem or progenitor cells in clinical trials has not been reported till date. The system AutoCulture® used in this study can automatically replace the culture medium, centrifuge cells, split cells, and take photographs for morphological assessment. We examined the feasibility of this system in a clinical setting.ResultsWe observed similar characteristics by both the culture methods in terms of the growth rate, gene expression profile, cell surface profile by fluorescence-activated cell sorting, surface glycan profile, and genomic DNA stability. These results indicate that AutoCulture® is a feasible method for the cultivation of human cells for regenerative medicine.ConclusionsAn automated cell-processing machine will play important roles in cell therapy and have widespread use from application in multicenter trials to provision of off-the-shelf cell products.