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Topical Gel of Mesenchymal Stem Cell-Conditioned Medium-induced Serum Injury Accelerates Wound Healing in Skin Excision Tissue Berlian, Mukti Arja; Alif, Iffan; Subchan, Prasetyowati; Handoyo, Frigi Eko; Husain, Sofian Azalia; Husni Ahmad Sidiq; Arlinda, Dyken Dwi; Adityani, Resanti
International Journal of Cell and Biomedical Science Vol 1 No 1 (2022)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v1i1.12

Abstract

Introduction: Umbilical cord-derived mesenchymal stem cells (UC-MSCs) accelerating wound closure by increasing VEGF and PDGF level leading to re-epithelialization, cell infiltration, and angiogenesis. It has been found that MSC-conditioned medium (MSC-CM) can enhance migration of fibroblasts in scratch assays. However, the effect of MSC-CM-induced serum injury (MSC-CM-S) formulated in gel to accelerate wound healing remains unclear. This study aims to evaluate the effect of several doses of topical gel of MSC-CM-S in accelerating wound healing. Methods: The MSCs were cultured medium-supplemented serum injury of wounded rat (8:1) to get MSC-CM-S. The topical gel of MSC-CM-S was made by base gel supplemented with MSC-CM-S. Eighteen Wistar rats were randomly assigned into control (C) and treatment groups (T1, T2). Groups were received serum-free medium gel (C), 25 µl MSC-CM-S in topical gel (T1), 50 µl MSC-CM-S in topical gel (T2), twice daily for 9 days. PDGF and VEGF level and fibroblast density were measured by ELISA and HE staining at day 3 and 6, respectively. Results: This study showed that there was significant increase of VEGF and PDGF level along with a significant increase of fibroblast density at day 3 and 6. The T2 showed optimum enhancement level of VEGF, PDGF and fibroblast density. Conclusion: Topical gel of MSC-CM-S was effective to accelerate wound closure by enhancing PDGF and VEGF level in full-thickness skin defect rats.
The Effect of Hypoxia on the Soluble Molecules of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Widyatmoko, Agus; Alif, Iffan; Irawan, Risky Candra Satria; Handoyo, Frigi Eko; Sidiq, Husni Ahmad
International Journal of Cell and Biomedical Science Vol 1 No 3 (2022)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v1i3.24

Abstract

Background: Umbilical cord-derived stem cells (UC-MSCs) are essential cell sources for cell-based therapies in regenerative medicine. Hypoxia is a key element of the stem cell niche and crucial for the preservation of UC-MSCs properties. The normal growth conditions for UC-MSCs are under atmospheric oxygen concentrations of 20–21%. However, the administration of UC-MSCs in inflammatory conditions provides oxygen-deficient environments. Thus, treating UC-MSCs with low oxygen exposure provides them with more survival and recovery potential. Objective: In this study, we assessed the impact of hypoxia incubation for 12 h on the UC-MSCs proteome. Methods: UC-MSCs were isolated from UC patients regarding informed consent. At passage 5, in 80% confluent, UC-MSCs were incubated in 5% O2 for 12 h. The morphology of UC-MSCs was assessed using a microscope. The level of FGF-2, FGF-8, TNF-α, and HSP-70 were analyzed using ELISA. Results: Hypoxic condition could change their morphology and enhance the cellular density compared to normoxic conditions in vitro. The level of FGF-2, FGF-8, TNF-α, and HSP-70 were significantly increased after the hypoxic condition of UC-MSCs compared to normoxia. Conclusion: Our findings suggest that the hypoxic condition was able to induce survival capacity and soluble molecules secreted by UC-MSCs.
Priming of Mesenchymal Stem Cells for Enhanced Interleukin-10 Secretion via Conditioned Medium from Lipopolysaccharide-Activated Peripheral Blood Mononuclear Cells Prasetio, Ardi; Syafitri, Luthfiana Mifta; Prabowo, Adam; Alif, Iffan; Nurichsan, Aldan
International Journal of Cell and Biomedical Science Vol 2 No 5 (2023)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v2i5.39

Abstract

Background: Mesenchymal stem cells (MSCs) are known for their immunomodulatory properties, particularly their ability to secrete anti-inflammatory cytokines such as interleukin-10 (IL-10). Enhancing the secretion of IL-10 by MSCs could have significant therapeutic potential for treating inflammatory diseases. Objective: This study aimed to prime the secretion of IL-10 by MSCs through the use of conditioned medium (CM) derived from lipopolysaccharide (LPS)-induced peripheral blood mononuclear cells (PBMCs). Methods: MSCs were isolated from Wharton’s Jelly and characterized using flow cytometry and differentiation assays. PBMCs were isolated from human blood samples and stimulated with LPS to produce a pro-inflammatory environment. The conditioned medium from these LPS-induced PBMCs was collected and add to MSCs culture medium in 5% and 7.5%. After 24h and 48h incubation, IL-10 secretion by MSCs was measured using an enzyme-linked immunosorbent assay (ELISA). Results: The results demonstrated that MSCs cultured in the conditioned medium from LPS-induced PBMCs showed a significant increase in IL-10 secretion compared to control conditions in 24h exposure, but not significantly different in 48h. Conclusion: The exposure of conditioned medium from LPS-induced PBMCs may effectively enhances the secretion of IL-10 by MSCs.
Comparison of Two Tangential Flow Filtration Methods in Isolating CD63+/CD9+ Mesenchymal Stem Cell Exosome Putra, Agung; Alif, Iffan; Prasetio, Ardi; Prawitasari, Salindri
International Journal of Cell and Biomedical Science Vol 2 No 4 (2023)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Background: Extracellular vesicles, particularly CD63+/CD9+ Mesenchymal Stem Cell Exosome (MSC-Exo), have emerged as crucial mediators of intercellular communication and potential therapeutic agents, including regenerative medicine and immunomodulation. However, the precise isolation and purification of MSC exosomes pose critical challenges. Tangential Flow Filtration (TFF) has gained recognition as an efficient exosome isolation method, offering scalability and versatility. In this study, we address the pressing need for standardized exosome isolation methods by comparing two distinct TFF-based protocols for isolating CD63+/CD9+ MSC exosomes based on filter size pore order. Methods: MSC-Exo were conducted from the Stem Cell and Cancer Research Laboratory (SCCR Indonesia), which were then processed through TFF using different filter sizes and orders. There are two filtration methods compared, first, MSC-Exo was filtered with 1000-5-500-300-100-50-10-5 filter order. Second procedure, MSC-Exo was filtered using 1000-500-300-100-50-10-5 filter order. Result: Flow cytometry analysis revealed variations in the percentage of CD63+/CD9+ in the MSC-Exo based on filter order. The results indicate that the choice of filter order significantly influences the size range with the highest concentration of CD63+/CD9+ MSC-Exo. Conclusion: This research underscores the importance of optimizing TFF-based isolation methods for CD63+/CD9+ MSC exosomes, especially in the order of filter pore size.
Hypoxic MSCs Secretome Modulates IL-18-Mediated Inflammatory in Type 2 Diabetes Mellitus via AP-1 Regulation Amansyah, Fajar; Alif, Iffan; Irawan, Risky Candra Satria
International Journal of Cell and Biomedical Science Vol 3 No 7 (2024)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v3i7.47

Abstract

Background: Chronic inflammation is central to the pathophysiology of Type 2 Diabetes Mellitus (T2DM), contributing to the progression of metabolic dysfunction characterized by hyperglycaemia and insulin resistance. This study aims to investigate the therapeutic potential of the hypoxic MSCs secretome (SH-MSCs) in reducing inflammation of a T2DM rat model. Methods: T2DM was induced in Wistar rats through a high-fat diet (HFD) followed by streptozotocin (STZ) administration. A total of 24 healthy male Wistar rats were randomly assigned to five groups: healthy control, T2DM, T2DM + metformin, T2DM + SH-MSCs. Results: SH-MSCs significantly reduced IL-18 mRNA expression, a key indicator of proinflammation, and suppressed the expression of AP-1 mRNA, a crucial proinflammatory transcription factor. Conclusion: These findings highlight the therapeutic potential of SH-MSCs as an alternative approach to alleviate inflammation in T2DM.
Generation of M2c Macrophages Using IL-10 Exposure In Vitro Alif, Iffan; Kurniawan, Dicky Zulfa F; Wilaksono, Bhirau; Susanto, Indra
International Journal of Cell and Biomedical Science Vol 3 No 8 (2024)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v3i8.52

Abstract

Background : Macrophages are essential cells of the innate immune system that highly adaptable and play vital roles in tissue homeostasis, immune regulation, and development. Among their various phenotypes, the M2c subtype—induced by IL-10 and TGF-β—is known for its regulatory functions, high phagocytic capacity, and pro-angiogenic potential. This study aimed to investigate whether M2c polarization via IL-10 stimulation occurs in a dose-dependent manner by differentiating human monocyte-derived macrophages using IL-10 at concentrations of 5, 10, and 20 ng/mL. Methods : IL-10 expression levels were assessed by ELISA and qPCR. The results demonstrated a clear dose-dependent increase in IL-10 expression across both methods. Results : ELISA measurements showed IL-10 levels increasing from a mean of 3.2±0.7 pg/mL in untreated controls to 19.34±2.3 pg/mL (5 ng/mL), 35.5±7 pg/mL (10 ng/mL), and 67.2±5.8 pg/mL (20 ng/mL). Similarly, qPCR analysis revealed a corresponding increase in IL-10 gene expression, with relative fold changes from 1±0 (untreated control) to 3.1±0.4, 6±1.3, and 10.1±1.6, respectively. Conclusions : These findings indicate that IL-10 induces M2c macrophage polarization in a dose-dependent manner, providing insight into optimized conditions for generating regulatory macrophage populations for potential therapeutic applications.