Project description:Here, we chemically induced hepatocyte-like cells (CiHeps) from mouse embryonic fibroblasts (MEFs) using a new protocol. To illustrate the transcriptomic landscape of hepatocyte cell formation by chemical induction, scRNA-seq was performed in C6F5UE-C6F(L)V-induced MEFs at 60 days of induction. The analysis of scRNA-seq data revealed that gastrointestinal-like and keratinocyte-like cells were induced along with CiHeps and these induced epithelial cells closely resembled that of their primary counterparts.
Project description:Here, we chemically induced hepatocyte-like cells (CiHeps) from mouse fibroblasts using a new protocol. To compare the transcriptional profiles between different cocktail-induced cells and primary hepatocytes (pHeps), we profiled the transcriptomes of induced cells and freshly isolated primary mouse hepatocytes. CiHeps' function and maturation-related gene expression was validated as well as the similarity it shares with pHeps.
Project description:Here we report the reprogramming of human fibroblasts to produce chemically-induced osteogenic cells (ciOG), and explore the potential uses of ciOG in bone repair and disease treatment. A chemical cocktail of RepSox, forskolin, and phenamil was used for osteogenic induction of fibroblasts by activation of RUNX2 expression. Following a maturation, the cells differentiated toward an osteoblast phenotype that produced mineralized tissue. Bulk RNA sequencing revealed that up- and down-regulated genes in ciOG relative to aHDF resembled those in hOB during maturation.
Project description:Cardiovascular progenitor cells (CPCs) can be reprogrammed from somatic fibroblasts by combinations of genes, providing a new avenue for cardiac regenerative therapy. Here we developed a strategy to capture chemically induced CPCs (ciCPCs) during the small molecule-induced cardiac reprogramming of mouse fibroblasts and expand these ciCPCs for a long-term in a chemically defined condition. RNA-seq analysis has been used to compare the expression profile of ciCPCs at early and late passages.
Project description:Recently, direct reprogramming between divergent lineages has been achieved by introducing cell-fate-determining transcription factors. This progress may provide alternative cell resources for drug discovery and regenerative medicine. However, the genetic manipulation may limit the future application of these approaches. In this study, we identified a novel small-molecule cocktail that directly converted fibroblasts into neuronal cell fate with a high yield up after 16-days of induction. After a further maturation stage, these chemically-induced neurons (CiNs) possessed neuron-specific expression patterns, generated action potentials and formed functional synapses. Gene expression profiling revealed the activation of neuronal specific genes in the early stage of small molecule treatment. Overall, our findings prove the principle of chemically-induced direct reprogramming of somatic cell fates across germ layers without genetic manipulation, and show that cell fate can be manipulated through disrupting initial cell program and activating target cell master genes with pure chemicals. Total of 15 samples were analyzed, including mouse fibroblasts, mouse cortical primary neurons and chemically-induced neurons by different duration of chemical induction (Day0, Day4, Day8, Day19) and different small-molecule cocktail (FICB, FICB-1)
Project description:Plasticity of differentiated cells has been proved by nuclear transfer, induced pluripotent cells and transdifferentiation. Here we show that by transduction of 3 factors (FOXA3, HNF1A and HNF4A), human fetal fibroblasts can be converted to hepatocyte-like cells (hiHep cells), expressing hepatic marker genes, and acquiring many mature hepatocyte functions in vitro and in vivo. Human fetal fibroblasts (HFF) were tranfected with 3 liver enriched transcription factors (FOXA3, HNF1A, HNF4A), and converted to hepatocyte-like cells (hiHep cells). HFF and primary human hepatocytes (PHH) serve as control.