Abstract
Background: The senescence process in human dermal fibroblasts (HDFs) is caused by cell cycle withdrawal processes, one of which is the result of the retinoblastoma (Rb) protein being in a hypo-phosphorylated state. Since adipose-derived stem cells (ADSCs) have a paracrine effect, ADSCs were utilized to improve the senescence process of HDFs. The use of non-autologous cell culture media to grow ADSCs can be legally problematic; therefore, platelet-rich plasma (PRP) can be considered as an alternative medium. PRP contains various growth factors that can be used to process the reversal of senescent HDFs. The combination of ADSCs and PRP is expected to increase the expression of Rb protein in HDFs that have undergone the senescence process. Methods: This study was performed in vitro with a randomized sample, and non-blinded pre-and post-test control group. The primary culture of senescent HDFs was transfected with a combination of ADSCs and 10% PRP. The effect on migration was observed through the scratch test, while the effect of PRP on reversal senescence was observed through Sa-β-gal analysis and the expression of protein Rb with ELISA. Results: The senescent HDFs that received a combined transfection of ADSCs and 10% PRP proliferated rapidly in the scratch test. Based on the Sa-β-gal assay, they showed fewer senescent HDFs cells. The combination of ADSCs and 10% PRP elevated the expression of Rb protein significantly (P < 0.001). Conclusions: The combination of ADSCs and 10% PRP was shown to have a reversal effect on the senescence process of HDFs in vitro.
Highlights
Adipose-derived stem cells (ADSCs) are mesenchymal stem cells that reside in fat tissue and were first identified by Zuk et al.[1]
The human dermal fibroblasts (HDFs) media was removed and the cells were transfected with the adipose-derived stem cells (ADSCs) - 10% platelet-rich plasma (PRP) for another 72 h and subsequently examined by enzyme-linked immunosorbent assay kit (ELISA) to determine the Rb protein level
The scratch test showed that the senescent HDF cell group that received the ADSCs – 10% PRP medium proliferated faster than the control group (Figure 1)
Summary
Adipose-derived stem cells (ADSCs) are mesenchymal stem cells that reside in fat tissue and were first identified by Zuk et al.[1]. FBS is important in the cell culture process When it is used for regenerative medicine, this serum is less safe because it contains xenoproteins, which can cause rejection or transmission of infectious products. Nowadays, the use of non-autologous cell culture media, like FBS in ADSCs for clinical applications is legally challenged. Since adipose-derived stem cells (ADSCs) have a paracrine effect, ADSCs were utilized to improve the senescence process of HDFs. The use of non-autologous cell culture media to grow ADSCs can be legally problematic; platelet-rich plasma (PRP) can be considered as an alternative medium. Results: The senescent HDFs that received a combined transfection of ADSCs and 10% PRP proliferated rapidly in the scratch test. Conclusions: The combination of ADSCs and 10% PRP was shown to have a reversal effect on the senescence process of HDFs in vitro
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