Conditioned Media of Human Umbilical Cord Blood Mesenchymal Stem Cell Inhibits Ultraviolet B-induced Apoptosis in Fibroblasts

Abstract

BACKGROUND: Ultraviolet (UV)B irradiated-skin cells may respond either by activating protective or apoptotic mechanisms. Several treatments has been reported to prevent apoptosis of the skin cell. To date, despite of the cell, secretome of mesenchymal stem cell (MSC) has been progressively explored for various treatment. Current research was conducted to find out the possible effect of MSC's secretome to protect normal cells from the UVB irradiation.METHODS: For preparation of the conditioned media human umbilical cord blood mesenchymal stem cell (CM-hUCB-MSC), hUCB was collected and separated to collect mononuclear cell (MNC). MNC was cultured in MSC medium until passage 4, then checked for MSC biomarkers. Conditioned media was then produced from the cultured MSC. For induction of apoptosis, NIH3T3 cells were pretreated with/without CM-hUCB-MSC, prior to the UVB irradiation for 5-60 minutes. After 24 hours, apoptosis evaluation was conducted with Sub-G1 assay with hypotonic fluorochrome solution using flow cytometer.RESULTS: The average of apoptotic cells irradiated with UVB for 30 minutes was 94.2%. The 30-minutes-UVB irradiation significantly induced apoptosis in NIH3T3 cells (p=0.000). The averages of apoptotic cells by pretreatment of 0, 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation were 94.2, 46.8, 31.8 and 31.5 %, respectively. The pretreatment of 5, 10 and 20% CM-hUCB-MSC prior to UVB irradiation could significantly decrease the percentage of apoptosis caused by UVB (p=0.001).CONCLUSION: Taken together, CM-hUCB-MSC inhibited UVB-induced apoptosis in NIH3T3 cells significantly, suggesting that CM-hUCB-MSC might be a potential anti-UVB-induced apoptosis factor. Hence, further research should be explored to disclose its specific intracellular mechanism.KEYWORDS: UVB, stem cell, conditioned media, fibroblast, apoptosis, aging, secretome, NIH3T3 cell

Mesenchymal Stem Cell Secretome for Dermatology Application: A Review

“…58 UV radiation can also cause increased levels of MMP1 and MMP9, resulting in inhibited synthesis of procollagens and triggering activation of various signaling pathways, namely mitogen activated protein kinase (MAPK), protein activator 1 (AP-1) and nuclear factor kappa B (NF-κB), which contributes to cell damage due to ROS formation and can trigger the apoptosis of skin cells, so as to inhibit skin regeneration and be able to inflict DNA damage in keratinocytes. 24,25,28,29,77 Various studies have been conducted to investigate the effects of secretome as an agent in the fight against photoaging. In vitro testing was done by various methods.…”

“…Methods that were used looked at cell proliferation activity, cellular aging and cellular apoptosis, intracellular ROS levels, and pyrimidine cyclobutane dimer assay (CPD). 26,27,29,58,78 Wang et al reported that there was an increase in cell proliferation from adipose-derived stem cells (ADSC-CM) use, as well as a decrease in the number of cells undergoing apoptosis and a decrease in cellular aging in HDFs. 58 Kim et al also found ADSC-CM can decrease the number of cell deaths from apoptosis (characterized by a decrease in the sub-G1 phase) and increase collagen I synthesis, as well as inhibit the production of MMP1 in HDFs.…”

“…60 Using the same method of sub-GI assay, Dewi et al also reported a reduction in the percentage of apoptosis after treatment with the human umbilical cord blood mesenchymal stem cell (hUCB-MSC-CM) conformation 5%, 10% and 20% from 94.2% to 46.8%, 31.8% and 31.5%, respectively. 29 The use of secretome was able to reduce intracellular ROS levels using the DCFDA fluorescent assay, 26 as well as able to increase cell migration and reduce the level of formation of CPD in vitro. 27 The secretome of ADSCs had the effect of providing photoprotection.…”

Coffea canephora Bean Extract Induces NIH3T3 Cell Migration

“…(1) Wound healing is crucial after tooth extraction. Wound healing can be induced with several agents, including cells (2) and their factors (3). In wound healing process, fibroblasts are required as the source of the tissue.…”