Stem Cells and Cloning-Advances and Applications最新文献

Introduction: The therapeutic application of adipose-derived stem cells (ASCs) for wound healing has been reported. However, consistently controlling ASC secretory-factor levels, scaling up, and their mechanism of action have been poorly understood, which are critical steps for developing biological drugs to treat second-degree burns.

Objective: Our goal is to develop a biological product, named consortia factors-δ2 (CFx-δ2), derived from cell-to-cell interactions between human immortalized ASCs using our patented technology and to evaluate the product consistency and in vitro biological effects to enhance the wound healing process.

Methods: To assess product consistency, three batches of CFx-δ2 were analyzed and compared using immunomicroarray and metabolomics. The biological effects of these batches were studied using an in vitro wound healing assay and ex vivo human skin explants subjected to burn wounds. Anti-inflammatory effects were analyzed by inhibition of Nuclear Factor- κB (NF-κB) nuclear translocation in human dermal endothelial cells (HMEC-1). Finally, pro-angiogenesis was evaluated by the induction of tube-like structures and capillary networks in HMEC-1 cells promoted by CFx-δ2.

Results: Our technology enabled the production of CFx-δ2 with enhanced efficiency and quality. When the three batches were compared, we found high consistency in the pattern and levels of cytokines as well as a strong correlation in the metabolite pattern. Accelerated wound healing was observed after in vitro and ex vivo wound treatment with CFx-δ2 in comparison to controls. The nuclear translocation of NF-κB, induced by Lipopolysaccharides (LPS), was 50% inhibited by CFx-δ2. Induction of more tube-like structures and networks was observed in endothelial cells treated with CFx-δ2 in comparison to controls.

Discussion: These findings demonstrate that our technology is efficient to derive and scale up consistent levels of CFx-δ2 from immortalized adipose stem cells. The product accelerates wound healing by increasing fibroblast proliferation and migration, inhibiting inflammation, and promoting angiogenesis.

Introduction: Mesenchymal stem cells (MSCs) have a paracrine impact and may regenerate a variety of tissues. This represents a new prospect in cell-based stroke treatment. Several in vitro and in vivo investigations have demonstrated the neuroprotective and neurogenesis properties of MSCs and their secretome.

Purpose: This review provides a comprehensive analysis of the therapeutic effects of MSCs and their secretome on stroke models in vitro and in vivo.

Methods: A coverage evaluation is undertaken in accordance with PRISMA-ScR principles. The selection procedure includes the identification of items. Scopus site, PubMed and ScienceDirect, are used for in vitro and in vitro research, including electronic searches. The search terms include "ischemic stroke" or "MCAO", "MSC", "secretome", and "neurogenesis" or "angiogenesis". The searches are limited to English-language articles with full text availability.

Results: After selecting 390 papers from two search engines, 94 publications satisfied the review criteria for using MSCs and secretomes for ischemic stroke treatment. We comprehensively review both in vitro and in vivo studies, analyzing aspects such as the source and treatment of MSCs and secretomes, as well as administration, dosage, and mechanisms of therapeutic effects in stroke models.

Conclusion: MSC and secretome therapy for stroke have shown promising results in both in vitro and in vivo models. Exploration of alternative MSC sources, refining of isolation techniques, transfection of various proteins, and combination with herbal medicine are all efforts to improve the preclinical model. This work can be used as a reference for preclinical researchers to help with research design and translational research in clinical trials.

Background: Chronic tendon injuries often lead to diminished healing capacity, necessitating innovative treatments. Mesenchymal stem cells (MSCs) secretome has emerged as a promising option for enhancing tendon repair through paracrine signaling. This study evaluates the effectiveness of MSC secretome, derived from tendon-derived stem cells (TDSCs) and adipose-derived stem cells (ASCs) in healing chronic Achilles tendon injuries in a rat model. The focus is on Procollagen Type I N-Terminal Peptide (PINP) and Procollagen Type III N-Terminal Peptide (PIIINP) levels, and histopathological changes.

Methods: Fourteen adult male rats were divided into four groups: Group I (TDSC secretome), Group II (ASC secretome), Group III (combination of TDSC and ASC secretome), and Group IV (control). The healing response was assessed through PINP and PIIINP immunoserological markers, and histopathological changes were analyzed. The study adhered to ARRIVE and ICLAS guidelines and followed the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals.

Results: The combination group showed significantly higher PINP levels compared to the control group (p = 0.004), suggesting enhanced Type I collagen synthesis. However, no significant differences in PIIINP levels were observed among the groups. Histopathological analysis showed no significant differences in collagen alignment or angiogenesis between treatment and control groups.

Conclusion: The MSC secretome, particularly the combination of TDSCs and ASCs, may accelerate collagen Type I synthesis and improve tendon microstructure. This suggests their potential for treating chronic tendon injuries. However, further research with longer observation periods and clinical trials is crucial to confirm these findings and advance our understanding of tendon healing.

The mammalian oocyte is pivotal in reproductive biology, acting as a central hub for cellular reprogramming and stemness. It uniquely contributes half of the zygotic nuclear genome and the entirety of the mitochondrial genome, ensuring individual development and health. Oocyte-mediated reprogramming, exemplified by nuclear transfer, resets somatic cell identity to achieve pluripotency and has transformative potential in regenerative medicine. This process is critical for understanding cellular differentiation, improving assisted reproductive technologies, and advancing cloning and stem cell research. During fertilization, the maternal-zygotic transition shifts developmental control from maternal factors to zygotic genome activation, establishing totipotency. Oocytes also harbor reprogramming factors that guide nuclear remodeling, epigenetic modifications, and metabolic reprogramming, enabling early embryogenesis. Structures like mitochondria, lipid droplets, and cytoplasmic lattices contribute to energy production, molecular regulation, and cellular organization. Recent insights into oocyte components, such as ooplasmic nanovesicles and endolysosomal vesicular assemblies (ELVAS), highlight their roles in maintaining cellular homeostasis, protein synthesis, and reprogramming efficiency. By unraveling the reprogramming mechanisms inherent in oocytes, we advance our understanding of cloning, cell differentiation, and stem cell therapy, highlighting their valuable significance in developmental biology and regenerative medicine.

Objective: Chronic wounds are a common clinical problem that necessitate the exploration of novel regenerative therapies. We report a method to investigate the in vitro wound healing capacity of an innovative biomaterial, which is based on amniotic membrane-derived stem cells (AMSCs) embedded in an alginate hydrogel matrix. The aim of this study was to prepare an sodium alginate-based hydrogel, cross-linked calcium chloride (CaCl₂₎ with the active ingredient AMSC (AMSC/Alg-H) and to evaluate its in vitro effectiveness for wound closure.

Methods: This hydrogel preparation involved combining sterile solutions of AMSC, sodium alginate, and CaCl_2, followed by rinsing with serum-free media. The cells were cultured in different 6-well plates, namely sodium alginate, calcium chloride, AMSC, Alg-H, and AMSC/Alg-H, in complete medium with 10% FBS. The hydrogel was successfully formulated, as confirmed by characterization techniques including Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FTIR) spectroscopy, Differential Scanning Calorimetry (DSC), Cytotoxicity Studies, TGF-β1 Level Measurement by ELISA, and Cell Scratch Wound Assay.

Results: Cryo-EM characterization of the Alg-H preparation successfully demonstrated the encapsulation of MSCs. FTIR and DSC analyses indicate that crosslinking transpires in Alg-H encapsulating AMSC. The AMSC/Alg-H preparation showed no significant difference in toxicity compared to HaCaT cells (p < 0.05), indicating it was not toxic to HaCaT cells. Furthermore, in the scratch wound assay test at 24 hours, the AMSC/Alg-H preparation achieved 100% wound closure, outperforming both AMSC and Alg-H alone. In vitro assessment revealed that AMSC/Alg-H significantly enhanced key wound healing processes, including cell proliferation and migration, compared to Alg-H.

Conclusion: Our study demonstrated the promising potential of AMSC/Alg-H as an enhanced regenerative therapy for in vitro wound healing. AMSC/Alg-H was able to maintain the viability of AMSCs and facilitate the formation of tissue-like structures.

Biobanking has emerged as a transformative concept in advancing the medical field, particularly with the exponential growth of umbilical cord (UC) biobanking in recent decades. UC blood and tissue provide a rich source of primitive hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) for clinical transplantation, offering distinct advantages over alternative adult stem cell sources. However, to fully realize the therapeutic potential of UC-derived stem cells and establish a comprehensive global UC-biobanking network, it is imperative to optimize and standardize UC processing, cryopreservation methods, quality control protocols, and regulatory frameworks, alongside developing effective consent provisions. This review aims to comprehensively explore recent advancements in UC biobanking, focusing on the establishment of rigorous safety and quality control procedures, the standardization of biobanking operations, and the optimization and automation of UC processing and cryopreservation techniques. Additionally, the review examines the expanded clinical applications of UC stem cells, addresses the challenges associated with umbilical cord biobanking and UC-derived stem cell therapies, and discusses the promising role of artificial intelligence (AI) in enhancing various operational aspects of biobanking, streamlining data processing, and improving data analysis accuracy while ensuring compliance with safety and quality standards. By addressing these critical areas, this review seeks to provide insights into the future direction of UC biobanking and its potential to significantly impact regenerative medicine.

Background and objective: Non-obstructive azoospermia (NOA) is an important cause of male infertility. This study is being proposed to assess the efficacy of autologous bone marrow-derived mesenchymal stem cells (MSCs) in the reversal of busulfan-induced NOA in rats.

Methods: Twenty adult 3-month-old male rats were divided into two groups: a control group and a study group. In the study group, bone marrow was aspirated to culture MSCs. NOA was created by stopping endogenous spermatogenesis in all the animals by injecting two doses of busulfan 10 mg/kg body weight with a 3 week interval. Four weeks after the last dose of busulfan, two animals were euthanized and the testes were studied histologically to confirm complete azoospermia. In the study group, five million MSCs in 1 mL normal saline were injected into seminiferous tubules; and in the control group, 1 mL of normal saline was injected. After 4 weeks of MSC injection, all the rats were euthanized and epididymis tails and testes were harvested and sent for measurement of serological indices, including luminal, cellular, and total diameters, luminal, cellular, and cross-sectional areas, number of tubules per unit area of testis, numerical density of the tubules, and spermatogenesis index, pre- and post-MSC transplantation.

Results: The effect of busulfan on the testicular tissue was universally devastating. In the control group, there was variable length and width of markedly necrotic seminiferous tubules, whereas in the group treated with autologous bone marrow-derived MSCs there was variable height of germinal epithelium in seminiferous tubules, with active spermatogenesis, showing spermatogonia, spermatocytes, and sperm.

Conclusion: MSC injection in the testis has the potential to reverse the testicular function of spermatogenesis after cytotoxic therapy. Human trials should be undertaken to confirm our findings and bring the results into clinical practice.

Introduction: Mesenchymal stem/stromal cells (MSCs)-based products have unique characteristics compared to other drugs because of their inherently variable effects depending on culture conditions and microenvironment. In some cases, cells can be produced individually, one batch at a time, for personalized therapy. Therefore, it is very important to optimize both culture conditions and medium composition under Good Manufacturing Practice (GMP) standards. MSCs properties have been exploited as potential cell therapies in regenerative medicine. The main mechanism of their protective and regenerative effect is based on their secretory activity. Simultaneously, their secretome is highly variable and sensitive to any change in environmental conditions. Depending on the type of damage and the target application, it is desirable to enhance the secretion of therapeutic factors. Changes in the modulation of environmental conditions can affect survival, migration ability, and both proliferative and clonogenic potentials.

Materials and methods: This study cultured Wharton's jelly-derived MSCs (WJ-MSCs) in media with varying concentrations of human platelet lysate (hPL). Two groups were created: one with low hPL concentration and another with a high hPL concentration. The effects of these different hPL concentrations were analyzed by assessing mesenchymal phenotype retention, secretory activity, clonogenic potential, proliferation, and migration capabilities. Additionally, the secretion levels of key therapeutic factors, such as Hepatocyte Growth Factor (HGF), Brain-Derived Neurotrophic Factor (BDNF), and Chemokine Ligand 2 (CCL-2), were measured.

Results: WJ-MSCs maintained their mesenchymal phenotype regardless of hPL concentration. However, a higher concentration of hPL promoted cell clonogenic potential, proliferation, migration, and increased secretion of therapeutic factors.

Conclusion: Adjusting the hPL concentration in the culture medium modulates the response of WJ MSCs and enhances their therapeutic potential. Higher hPL concentration promotes increased secretory activity and improves the regenerative capacity of WJ-MSCs, suggesting a promising strategy to optimize MSC-based therapies.

Various studies have been widely conducted on conditioned medium for the development of anti-aging preparations, including the utilization of stem cells, which present a promising alternative solution. This narrative review aims to understand the latest developments in various conditioned medium stem cell applications for anti-aging on the skin. A search of the Scopus database yielded publications of interest. The research focused on articles published without restrictions on the year. After finding 68 articles in the search results, they moved on to the checking phase. Upon comprehensive literature review, 23 articles met the inclusion criteria, while 45 articles were deemed ineligible for participation in this research. The results of the review indicate that conditioned medium from various stem cells has demonstrated success in reducing risk factors for skin aging, as proven in various tests. The successful reduction of the risk of skin aging has been established in vitro, in vivo, and in clinical trials. Given the numerous studies on the progress of exploring and utilizing conditioned medium, it is expected to provide a solution to the problem of skin aging.

[This retracts the article DOI: 10.2147/SCCAA.S212087.].