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Downregulation of IL-6 and TNF-α Expression with Mesenchymal Stem Cells Therapy in Allergic Rhinitis Rats Models Restimulia, Lia; Dewi, Dian Andriani Ratna; Nazar, Mohammad Ariq; Irawan, Risky Chandra Satria; Ghaissani, Shabrina Syifa; Haryono, Erlina
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.v1i1.13

Abstract

Rhinitis is an inflammatory that characterized by nasal symptoms due to the condition of the nasal mucosa that trigerred by an interaction between environmental allergens and immunoglobulin (Ig)E. It is driven by host factors, infection, pathogens and various inflammatory pathways such as TNF-α that released in allergic responses from both mast cells and macrophages through IgE-dependent mechanisms. Secretome hypoxia mesenchymal stem cells (SH-MSCs) contain anti-inflammatory soluble molecules were able to improve the conditions of endothelial damage, inflammation and oxidative stress by decreasing levels of IL-6, TNF-α, ROS and increasing eNOS. This study will investigate the effect of SH-MSCs at a dose of 300 µL on TNF- and IL-6 expression in rat model ovalbumin-induced rhinitis in vivo. Thirty male Wistar rats were randomly divided into 3 groups: control, treatment and sham group that administrated through intramuscular injection. The results of this study found that SH-MSCs can downregulate the inflammatory cytokines TNF-α and IL-6 in rat model ovalbumin-induced rhinitis in vivo
Surface Marker Expression and Morphological Alterations in Umbilical Cord-Derived MSCs Over Passages 4 to 9: A Flow Cytometry and Microscopic Analysis Prawitasari, Salindri; Ayu, Dian Respati; Irawan, Risky Chandra Satria; Prabowo, Adam
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.v3i9.51

Abstract

Background : Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are promising candidates for regenerative medicine due to their high proliferative capacity, multilineage differentiation potential, and low immunogenicity. However, prolonged in vitro expansion may lead to phenotypic drift and morphological changes that could impact their therapeutic efficacy. This study aimed to evaluate the expression of mesenchymal surface markers (CD73, CD90, CD105) and morphological characteristics of UC-MSCs from passage 4 (P4) to passage 9 (P9). Methods: Flow cytometry was employed using the BD Stemflow™ Human MSC Analysis Kit to quantify the expression of both positive and negative surface markers, while morphological assessments were performed via phase-contrast microscopy. Results : The results revealed that UC-MSCs maintained high expression levels of CD73, CD90, and CD105 across all passages, although a slight decline was observed in later passages. Morphological analysis indicated a transition from spindle-shaped, fibroblast-like cells at early passages to more enlarged and flattened cells with signs of senescence at higher passages. Conclusions : These findings suggest that although UC-MSCs retain their phenotypic identity up to P9, subtle morphological and marker expression changes may occur, underscoring the importance of passage selection in therapeutic applications. This study contributes to the optimization of UC-MSC culture protocols for standardized and effective clinical use.
Hypoxia-Induced Modulation of OCT-4, SOX-2, NANOG, and KLF-4 Expression in Mesenchymal Stem Cells Irawan, Risky Chandra Satria; Hidayah, Nurul; Wilaksono, Bhirau
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.v3i9.62

Abstract

Background: Mesenchymal stem cells (MSCs) possess remarkable regenerative and immunomodulatory capabilities, largely attributed to their intrinsic stemness properties regulated by transcription factors such as OCT-4, SOX-2, NANOG, and KLF-4. However, in vitro culture under normoxia often leads to stemness decline. Hypoxic preconditioning has been proposed to preserve stemness, though the optimal duration of exposure remains unclear. This study aimed to evaluate the time-dependent effects of hypoxia on the expression of core stemness-related transcription factors in human umbilical cord Wharton’s jelly-derived MSCs. Methods: MSCs (passage 5) obtained from the Stem Cell and Cancer Research (SCCR) Laboratory were cultured under hypoxia (5% O₂) for 0, 4, 6, 10, 12, and 24 hours. Gene expression of OCT-4, SOX-2, NANOG, and KLF-4 was quantified by qRT-PCR using GAPDH as a reference. Morphological changes were observed via phase-contrast microscopy. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Results: Hypoxic exposure induced a significant, time-dependent upregulation of OCT-4, SOX-2, and NANOG, with peak expression at 12 hours (approximately 13-, 7-, and 5-fold increases, respectively; p < 0.05). KLF-4 expression showed a modest elevation but was not statistically significant. Prolonged hypoxia (24 hours) resulted in a marked decline in all gene expressions toward baseline levels, accompanied by cell rounding and detachment. These findings indicate that short-term hypoxia enhances MSC stemness, while extended exposure triggers stress-related responses. Conclusion: Controlled short-term hypoxia (approximately 12 hours at 5% O₂) optimally enhances MSC stemness gene expression without compromising morphology or viability. This duration represents an effective preconditioning window for maintaining MSC multipotency and may improve the consistency and therapeutic efficacy of MSC-based applications.
Comparison of Animal Serum-Free Media Culture for cGMP-Compliant Mesenchymal Stem Cell Expansion Irawan, Risky Chandra Satria; Zakiyah, Nibras; Aji, Bramassetyo; Budiarto, Bagus Aji
International Journal of Cell and Biomedical Science Vol 4 No 11 (2025)
Publisher : Stem Cell and Cancer Research (SCCR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59278/cbs.v4i11.74

Abstract

Background: Mesenchymal stem cells (MSCs) are multipotent stromal cells distinguished by their ability to self-renew, their potential for multilineage differentiation, and their strong immunomodulatory characteristics. The recent in vitro expansion of MSCs primarily relies on culture media that is supplemented with fetal bovine serum (FBS). Although FBS is effective in enhancing the proliferation of MSCs and ensuring their survival, its use poses several notable challenges, particularly in clinical environments. Methods: This study seeks to fill this important gap by conducting a comprehensive comparison of various commercially available animal source-serum free media formulations against traditional FBS-supplemented media for the expansion of umbilical cord-derived mesenchymal stem cells (UC-MSCs). The morphology and density of the cells were examined using an inverted microscope. The cell surface of UC-MSCs was analysed through flow cytometry. The evaluation of cytokines released by UC-MSCs was carried out using ELISA. Result: The conditioned medium obtained from UC-MSCs cultured in HPL shows increased levels of exosomes and anti-inflammatory cytokines. Moreover, the UC-MSCs cultured in HPL-supplemented medium maintained a normal morphology and exhibited expression of UC-MSC surface markers exceeding 95%. Additionally, HPL enhanced the proliferation of UC-MSCs to eight times the cell number on the day of seeding. Conclusion: Media enriched with HPL presents considerable potential for future applications within the pharmaceutical sector.