ABSTRACT
The generation of functional retinal pigment epithelium (RPE) is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration (AMD). Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Best1::GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Best1::GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmentation upon activation of Retinoic acid (RA) and Sonic Hedgehog (SHH) signaling pathways. Our study not only established an ideal platform to investigate the transcriptional network regulating the RPE cell fate determination, but also provided an alternative strategy to generate functional RPE cells that complement the use of pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.
Subject(s)
Animals , Humans , Mice , Bestrophins , Cell Differentiation , Cell Line , Cell Lineage , Chloride Channels , Genetics , Metabolism , Embryonic Stem Cells , Cell Biology , Metabolism , Eye Proteins , Genetics , Metabolism , Fibroblasts , Cell Biology , Metabolism , Genes, Reporter , Green Fluorescent Proteins , Genetics , Metabolism , Pigmentation , Retinal Pigment Epithelium , Cell Biology , Metabolism , Transcription Factors , MetabolismABSTRACT
Objective To investigate the accuracy of the plasma DNA concentration in evaluating the prognosis in patients with sepsis.Methods One hundred and sixty patients with sepsis were enrolled as the sepsis group (group SE).Another 109 patients without sepsis hospitalized during the same period served as the control group (group C).The venous blood sample was taken on admission for determination of plasma DNA concentration by polymerase chain reaction,C reactive protein (CRP) concentration by ELISA.APACHE Ⅱ score and SOFA score were evaluated at 24 h after admission.The 160 patients with sepsis were divided into two groups according to the result of prognosis:survival group ( n =103) and death group ( n =57).Results Compared with group C,the plasma DNA concentration,CRP concentration,APACHE Ⅱ score and SOFA score were significantly increased in group SE (P＜0.05).Compared with survival group,the plasma DNA concentration,APACHEⅡ score and SOFA score were significantly increased in death group ( P ＜ 0.05).The areas under receiver operating characteristic (ROC) curves of the plasma DNA concentration was significantly larger than those of APACHE Ⅱ score and SOFA score (0.81(95％ CI,0.74-0.88) versus 0.68(95％ CI,0.60-0.77),or 0.72(95％ CI,0.63-0.82)).Conclusion The plasma DNA concentration can accurately evaluate the prognosis in patients with sepsis.As compared with the plasma CRP concentration,APACHE Ⅱ score and SOFA score,the plasma DNA concentration is more accurate to evaluate the prognosis in patients with sepsis.