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All Journal The Avicenna Medical Journal International Journal of Cell and Biomedical Science
Nugraha, Dendi Krisna
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Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Medicine & Pharmacology Public Health

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In Vitro Transcription as a Strategy to Enhance Mesenchymal Stem Cell Secretome for Therapeutic Use: An Overview Nugraha, Dendi Krisna; Falah, Ariq
International Journal of Cell and Biomedical Science Vol 3 No 9 (2024)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v3i9.61

Abstract

Mesenchymal stem cells (MSCs) have become a cornerstone of regenerative medicine owing to their capacity to secrete a diverse array of bioactive molecules, collectively termed the secretome. The MSC secretome exerts profound immunomodulatory, anti-inflammatory, and trophic effects that underpin much of the therapeutic efficacy observed in MSC-based interventions. Nevertheless, variability in the secretory profile across donors, tissue sources, and culture conditions continues to limit the reproducibility and potency of MSC-derived therapies. Recent advancements in in vitro transcription (IVT) mRNA technology have emerged as a robust and transient platform for the reprogramming of mesenchymal stem cells (MSCs) without the need for genomic integration. Through the use of IVT-mRNA-mediated expression of selected cytokines, growth factors, or homing receptors, MSCs can be endowed with enhanced anti-inflammatory and regenerative capabilities while preserving their native phenotype and viability. This review summarizes current IVT-mRNA–based strategies for engineering the MSC secretome, with an emphasis on augmenting anti-inflammatory cytokines (e.g., IL-10, TSG-6) and growth factors (e.g., VEGF, HGF, FGF2). The review also examines how IVT-mRNA redefines the cellular secretory landscape, outlines key considerations in IVT-mRNA design and optimization, and discusses translational implications for both cell-based and cell-free therapeutic applications. Finally, it underscores persistent challenges, including transient transgene expression, innate immune activation, and delivery inefficiency, and contemplates future prospects for integrating IVT-mRNA technology with advanced biomaterials, cellular priming methodologies, and multifactorial modulation to achieve consistent and potent therapeutic secretomes.
Human-Umbilical Cord-Mesenchymal Stem Cells (hUC-MCSs) Therapy with Extravesicles (EVs) Booster Improves Recovery in Type 2 Diabetes Mellitus with Cardiovascular Disease Nugraha, Dendi Krisna; Jutadi; Anggoro, Naufal Sebastian; Sari, Fikriya Novita; Ardani, Yanuar
International Journal of Cell and Biomedical Science Vol 4 No 10 (2025)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v4i10.66

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Background: Type 2 diabetes (T2DM) is a chronic metabolic disorder characterized by insulin resistance and β-cell dysfunction, leading to persistent hyperglycemia and complications. Studies have explored mesenchymal stem cell (MSC)-based therapies and their extracellular vesicles (EVs) as novel approaches for metabolic regulation and tissue repair. Case: A 43-year-old male patient exhibited symptoms including excessive thirst and hunger, frequent urination, fatigue, and intermittent blurry vision. He had type 2 diabetes and recently worsened symptoms. The obese patient had elevated blood glucose, HbA1c, triglycerides, and uric acid. He received umbilical cord-derived mesenchymal stem cells (161,6 × 106 cells), followed by seven intramuscular EV injections (1.5 cc each), along with diet and antioxidant supplements. Results: Three months after the conclusion of treatment, laboratory test showed significant improvement, with fasting glucose levels measuring at 91 mg/dL, HbA1c levels at 5,1%, triglyceride levels at 151 mg/dL, uric acid levels at 4,9 mg/dL, and an erythrocyte sedimentation rate of 12 mm/hr. The clinical symptoms such as nocturia, fatigue, and neuropathic pain, demonstrated a substantial improvement, as well as led to the resolution of skin xerosis and heel fissures. Conclusion: This case suggests that combined UC-MSC and EV therapy, complemented by lifestyle modification, may contribute to metabolic stabilization and symptomatic relief in T2DM patients.
Time-Dependent Secretion of IFN-γ, TNF-α, Perforin, and Granzyme-B in CTL-Conditioned Medium from Colorectal Cancer Patient Ibrahim, Sugeng; Putra, Agung; Hidayah, Nurul; Khan, Ahmed Faheem; Nugraha, Dendi Krisna
The Avicenna Medical Journal Vol. 6 No. 1 (2025): The Avicenna Medical Journal
Publisher : Faculty of Medicine, UIN Syarif Hidayatullah Jakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15408/avicenna.v6i1.46889

Abstract

Background: Cytotoxic T lymphocytes (CTLs) are key effectors of adaptive immunity, exerting their functions through the release of pro-inflammatory cytokines and cytotoxic molecules. Conditioned medium (CM) derived from CTLs has emerged as a potential cell-free immunotherapeutic strategy; however, the temporal dynamics of its secreted mediators remain poorly defined. Methods: Human CTLs were isolated, activated, and cultured in vitro. CM was harvested at defined intervals (days 5, 10, and 15). Concentrations of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), perforin, and granzyme-B were quantified by enzyme-linked immunosorbent assay (ELISA). Temporal secretion profiles were evaluated using ANOVA with post-hoc analysis. Results: IFN-γ and TNF-α exhibited peak secretion at day 5, followed by a decline at later time points. In contrast, perforin and granzyme-B increased progressively, reaching maximal levels at day 15. All four mediators demonstrated significant time-dependent variation (p < 0.05). Conclusion: CTL-derived CM displays distinct time-dependent secretion patterns, with cytokines predominating during early activation and cytotoxic molecules dominating later phases. These findings underscore the importance of optimizing CM collection timing to maximize its immunomodulatory and therapeutic potential, and provide a rationale for further translational development of CTL-CM in immunotherapy.
Co-Authors Agung Putra Anggoro, Naufal Sebastian Ardani, Yanuar Falah, Ariq Ibrahim, Sugeng Jutadi Khan, Ahmed Faheem NURUL HIDAYAH Sari, Fikriya Novita