199 EFFECTS OF REPROGRAMMING-CONDITIONED MEDIUM ON ULTRAVIOLET RAY A–DAMAGED HUMAN DERMAL FIBROBLASTS

2017 ◽  
Vol 29 (1) ◽  
pp. 208
Author(s):  
J. Kang ◽  
S. G. Lee ◽  
J. H. Kang ◽  
S.-M. Park ◽  
S. Y. Heo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Conditioned Medium ◽  
Pluripotent Stem Cells ◽  
Annexin V ◽  
Hierarchical Cluster ◽  
Stem Cell Differentiation ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Ultraviolet Ray

Ultraviolet ray A (UVA) is an electromagnetic light with a long wavelength from the sun. The penetration of UVA deep into the human dermis causes changes in cells, such as DNA fragmentation, apoptosis, and senescence, eventually leading a decline of proliferation and wound-healing ability. These changes induced by UVA exposure are similar to those seen in the process of stem cell differentiation. We postulated that the condition that reverses cellular differentiation may alleviate the UVA-induced damage in skin cells. Human dermal fibroblasts (HDF) could be reprogrammed to induced pluripotent stem cells (iPSC). Conditioned medium (CM) was prepared during the process of iPSC reprogramming (referred to as Repro-CM). The UVA-irradiated HDF were cultured in Repro-CM for 24 h. In comparison with CM prepared from the culture of normal HDF and iPSC (referred to as HDF-CM and iPSC-CM, respectively), effects of Repro-CM on UVA-irradiated cells were investigated. Viability, wound-healing ability, apoptosis, and senescence of HDF were analysed by WST-1 assay, scratch assay, Annexin V assay, and senescence-associated β-galactosidase assay, respectively. Upon recovering from the UVA-induced damage, viability and wound-healing ability of HDF were significantly different (P < 0.05) among the treatments in the order of Repro-, HDF-, and iPSC-CM. In the same context, apoptosis and senescence were significantly different (P < 0.05) in the order of iPSC-, HDF-, and Repro-CM. Interestingly, iPSC-CM did not substantially ameliorate UVA-induced damage, suggesting that the conditions optimized to pluripotent stem cells may not be suitable for the recovery from damage in terminally differentiated cells, such as fibroblasts. The RNA-seq analysis was performed to assess the genome-wide transcriptional profile in the process of recovery. Repro- and HDF-CM were categorized more closely than iPSC-CM in hierarchical cluster analysis. In comparison with iPSC-CM, the up-regulated genes by Repro-CM treatment were related to regulation of cell proliferation and cell metabolism, whereas down-regulated genes were related to antiapoptosis and response to stimulation of chemical and organic substances. Overall, providing an environment of reprogramming, as shown by Repro-CM in the present study, may assist recovery of HDF from UVA-induced damage. The results of the study may be applicable in developing pharmaceuticals to treat aging and wrinkling of the skin caused by UVA irradiation.

scholarly journals Brief Report: Inhibition of miR-145 Enhances Reprogramming of Human Dermal Fibroblasts to Induced Pluripotent Stem Cells

Stem Cells ◽  
2015 ◽  
Vol 34 (1) ◽  
pp. 246-251 ◽  
Author(s):  
Tomas Barta ◽  
Lucie Peskova ◽  
Joseph Collin ◽  
David Montaner ◽  
Irina Neganova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Induced Pluripotent

scholarly journals Regenerative Effects of Wharton’s Jelly Stem Cells-Conditioned Medium in UVA-Irradiated Human Dermal Fibroblasts

2018 ◽  
Vol 5 (1) ◽  
pp. 45-50
Author(s):  
Yohanes Widodo Wirohadidjojo ◽  
Arief Budiyanto ◽  
Hardyanto Soebono
Keyword(s):  
Stem Cells ◽  
Conditioned Medium ◽  
Cellular Proliferation ◽  
Positive Control ◽  
Dermal Fibroblasts ◽  
Wharton’S Jelly ◽  
Cellular Migration ◽  
Human Dermal Fibroblasts ◽  
Collagen Deposition ◽  
Wharton's Jelly

Background: Ultraviolet A radiation (UVA) can photo-age skin by suppressing the proliferation, migration, and collagen deposition of human dermal fibroblasts (HDFs). This process occurs because UVA light can inhibit the gene expression of the TGF-β receptor in HDFs. Moreover, Wharton’s Jelly Stem Cells-Conditioned Medium (WJSC-CM) is hypothesized to release microvesicles that contain short m-RNA with regenerative properties. Objectives: This study aimed to determine the regenerative properties of WJSC-CM on UVA-Irradiated Human Dermal Fibroblasts (UVA-HDFs) Methods: Passaged fourth of  HDFs obtained from the foreskin of six (11- to 13-year-old) boys were repeatedly irradiated with a total of 10 J/cm2 UVA and treated with various concentrations of WJSC-CM. We used non-irradiated HDFs as positive control. After that, the consumption of TGF-β, cellular proliferation, cellular migration, and collagen deposition of each group were measured and compared. Results: Compared to the non-irradiated groups, the proliferation rates, migration rates, and collagen deposition of UVA-HDFs significantly decreased (p<0.05). WJSC-CM can improve the consumption of TGF-β, proliferation, and cellular migration of UVA-HDFs. However, WJSC-CM failed to improve the collagen deposition of UVA-HDFs (p>0.05). Conclusions: WJSC-CM has regenerative properties and is a candidate material for the treatment of prematurely ageing skin induced by UVA-irradiation.   Ethical permission: This experiment was permitted by the local ethical permission committee Ref:KE/FK/382/EC with permission letter dated 17-04-2013.

Non-modified RNA-Based Reprogramming of Human Dermal Fibroblasts into Induced Pluripotent Stem Cells

2021 ◽  
Author(s):  
Immacolata Belviso ◽  
Franca Di Meglio ◽  
Veronica Romano ◽  
Stefania Montagnani ◽  
Clotilde Castaldo
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Induced Pluripotent

scholarly journals Non pharmacological high-intensity ultrasound treatment of human dermal fibroblasts to accelerate wound healing

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jeong Yu Lee ◽  
Dae-Jin Min ◽  
Wanil Kim ◽  
Bum-Ho Bin ◽  
Kyuhan Kim ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Intensity ◽  
Mapk Pathway ◽  
Dermal Fibroblasts ◽  
Ultrasound Treatment ◽  
Type I ◽  
Human Dermal Fibroblasts ◽  
Short Term ◽  
High Intensity Ultrasound ◽  
Extracellular Matrix Components

AbstractInspired by the effectiveness of low-intensity ultrasound on tissue regeneration, we investigated the potential effect of short-term high-intensity ultrasound treatment for acceleration of wound healing in an in vitro wound model and dermal equivalent, both comprising human dermal fibroblasts. Short-term ultrasound of various amplitudes significantly increased the proliferation and migration of fibroblasts and subsequently increased the production of the extracellular matrix components fibronectin and collagen type I, both of which are important for wound healing and are secreted by fibroblasts. In addition, ultrasound treatment increased the contraction of a fibroblast-embedded three-dimensional collagen matrix, and the effect was synergistically increased in the presence of TGF-β. RNA-sequencing and bioinformatics analyses revealed changes in gene expression and p38 and ERK1/2 MAPK pathway activation in the ultrasound-stimulated fibroblasts. Our findings suggest that ultrasound as a mechanical stimulus can activate human dermal fibroblasts. Therefore, the activation of fibroblasts using ultrasound may improve the healing of various types of wounds and increase skin regeneration.

Conditioned medium from the human umbilical cord-mesenchymal stem cells stimulate the proliferation of human keratinocytes

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Sushmitha Sriramulu ◽  
Antara Banerjee ◽  
Ganesan Jothimani ◽  
Surajit Pathak
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Conditioned Medium ◽  
Umbilical Cord ◽  
Human Keratinocytes ◽  
Human Umbilical Cord ◽  
Hacat Cells ◽  
And Migration

AbstractObjectivesWound healing is a complex process with a sequence of restoring and inhibition events such as cell proliferation, differentiation, migration as well as adhesion. Mesenchymal stem cells (MSC) derived conditioned medium (CM) has potent therapeutic functions and promotes cell proliferation, anti-oxidant, immunosuppressive, and anti-apoptotic effects. The main aim of this research is to study the role of human umbilical cord-mesenchymal stem cells (UC-MSCs) derived CM in stimulating the proliferation of human keratinocytes (HaCaT).MethodsFirstly, MSC were isolated from human umbilical cords (UC) and the cells were then cultured in proliferative medium. We prepared and collected the CM after 72 h. Morphological changes were observed after the treatment of HaCaT cells with CM. To validate the findings, proliferation rate, clonal efficiency and also gene expression studies were performed.ResultsIncreased proliferation rate was observed and confirmed with the expression of Proliferating Cell Nuclear Antigen (PCNA) after treatment with HaCaT cells. Cell-cell strap formation was also observed when HaCaT cells were treated with CM for a period of 5–6 days which was confirmed by the increased expression of Collagen Type 1 Alpha 1 chain (Col1A1).ConclusionsOur results from present study depicts that the secretory components in the CM might play a significant role by interacting with keratinocytes to promote proliferation and migration. Thus, the CM stimulates cellular proliferation, epithelialization and migration of skin cells which might be the future promising application in wound healing.

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