Biology subjects

Andrianto, A.

Publications and source records attributed to Andrianto, A..

2 recordsLinked to original sources

bioRxiv2020

Expression of SSEA4 and TRA1-60 as Marker of Induced Pluripotent Stem Cells by Small Molecule Compound VC6TFZ on Peripheral Blood Mononuclear Cell

IntroductionIt is possible to induce pluripotent stem cells from somatic cells, offering an infinite cell resource with the potential for disease research and use in regenerative medicine. Due to ease of accessibility, minimum invasive treatment, and can be kept frozen, peripheral blood mononuclear cells (PBMC) were an attractive source cell. VC6TFZ, a small molecule compound, has been successfully reprogrammed from mouse fibroblast induced pluripotent stem cells (iPSCs). However, it has not been confirmed in humans. ObjectiveThe aim of this research is to determine whether the small molecule compound VC6TFZ can induced pluripotency of PBMC to generate iPSCs detected with expression of SSEA4 and TRA1-60. MethodsUsing the centrifugation gradient density process, mononuclear cells were separated from peripheral venous blood. Mononuclear cells were cultured for 6 days in the expansion medium. The cells were divided into four groups; group 1 (P1), which was not exposed to small molecules (control group) and groups 2-4 (P2-P4), the experimental groups, subjected to various dosages of the small molecule compound VC6TFZ (VPA, CHIR, Tranylcypromine, FSK, Dznep, and TTNPB). The induction of pluripotency using small molecule compound VC6TFZ was completed within 14 days, then for 7 days the medium shifted to 2i medium. iPSCs identification in based on colony morphology and pluripotent gene expression, SSEA4 and TRA1-60 marker, using immunocytochemistry. ResultsColonies appeared on reprogramming process in day 7th. These colonies had round, large, and cobble stone morphology like ESC. Gene expression of SSEA4 and TRA 1-60 increased statisticaly significant than control group (SSEA4 were P2 p=0.007; P3 p=0.001; P4 p=0.009 and TRA 1-60 were P2 p=0.002; P3 p=0.001; P4 p=0.001). ConclusionSmall molecule compound VC6TFZ could induced pluripotency of human PBMC to generate iPSCs. Pluripotxency marker gene expression, SSEA 4 and TRA 1-60, in the experimental group was statistically significantly higher than in the control group.

molecular biology↗

bioRxiv2020

The Role of Human Platelet-Rich Plasma to Enhance the Differentiation from Adipose derived Mesenchymal Stem Cells into Cardiomyocyte: An Experimental Study

BackgroundAdipose derived mesenchymal stem cells (AMSCs) offer great potential to differentiate into cardiomyocyte. However, the optimal method to maximize the proliferation and differentiation is challenging. Platelet rich plasma (PRP) which contains high levels of diverse growth factors that can stimulate stem cell proliferation and differentiation in the context of cardiac tissue regeneration ObjectiveTo analyze the effect of PRP administration on the AMSCs differentiation into cardiomyocyte and compare to the group without PRP administration. MethodsThis study is a true experimental randomized post-test design study. AMSCs were isolated from adipose tissues and cultured until 4 passages. The samples were divided into 3 groups, i.e. negative control (-MEM), positive control (differentiation medium), and treatment group (PRP). The assessment of GATA-4 marker expression was conducted using flowcytometry on the fifth day and cTnT was conducted using immunocytochemistry on the tenth day to determine the differentiation to cardiomyocyte. Data analysis was conducted using T-test and One-Way ANOVA on normally distributed data determined through Shapiro Wilk test. ResultsFlowcytometry on GATA-4 expression revealed significant improvement on PRP group compared to negative and positive controls (67.04 {+/-} 4.49 vs 58.15 {+/-} 1.23 p < 0.05; 67.04 {+/-} 4.49 vs 52.96 {+/-} 2.02 p < 0.05). This was supported by the results of immunocytochemistry on troponin expression which revealed significant improvement on PRP group compared to negative and positive controls (38.13 {+/-} 5.2 vs 10.73 {+/-} 2.39 p < 0.05; 38.13 {+/-} 5.2 vs 26.00 {+/-} 0.4 p < 0.05). This was concordant to the hypothesis which stated that there was an effect of PRP administration on AMSCs differentiation into cardiomyocyte. ConclusionPRP administration on AMSCs culture significantly improve the differentiation to cardiomyocyte measured by GATA-4 and cTnT expressions.

molecular biology↗