Project description:Adipose tissue-derived stromal vascular fraction (Ad-SVF) contains stem cells and rarely transdifferentiate into spontaneously beating cardiomyocyte-like cells (beating CMs). However, the factors that regulate the differentiation of Ad-SVF into the cardiac lineage is unknown. We developed a simple culture protocol under which the adult murine inguinal Ad-SVF reproducibly transdifferentiates into beating CMs without induction. We used microarray analyses to evaluate the global gene expressions of the beating CMs and investigated the mechanism underlying the transdifferentiation of SVF cells into CMs.
Project description:We performed bulk RNA sequencing of several subpopulations of adipose tissue stromal cells to better understand the function of preadipocyte subpopulation (P1 and P2) in mouse inguinal adipose tissue. P1 and P2 cells isolated from inguinal adipose tissue from male and female C57/B6 mice, collected by FACS. P1 cells are CD31-CD45-TER119-SCA1+CD55+VAP1-CD142- cell population, and P2 cells are CD31-CD45-TER119-SCA1+CD55-VAP1+CD142- cell population. 50,000 cells were collected for each populations.
Project description:Adipose-derived stromal/stem cells (ASC) capable of multipotential differentiation can be isolated with high yield from human subcutaneous lipoaspirates. This study reports our recent experience isolating and immunophenotypically characterizing ASCs from >60 human subjects
Project description:Background: Adipose tissue-derived stromal cells (ATSCs) hold great promises in regenerative medicine, due to their easy retrieval, high proliferative capacity, and multi-lineage differentiation potential. In the last decade, several studies have reported the plasticity of ATSCs toward a hepatic fate. Nonetheless, the molecular mechanisms underlying the conversion from a mesenchymal to an epithelial phenotype remain poorly understood. Aim: In this study, we compared the full genome expression profiles of ATSCs cultured for 4 weeks under pro-hepatogenic conditions to undifferentiated ATSCs, in order to depict the molecular events involved in ATSC hepatic transdifferentiation. Methods: Molecular analysis was performed using the Affymetrix human focus arrays. Sets of differentially expressed genes were functionally categorized in order to understand which pathways drive the hepatic conversion and interesting target genes were validated by Q-PCR. Results: We showed that ATSC-derived hepatocyte-like cells activate several genes associated with specific liver functions, including protein metabolism, innate immune response regulation, and biodegradation of toxic compounds. Furthermore, microarray analysis highlighted the downregulation of several transcripts involved in stemness maintenance along with genes associated with a mesenchymal phenotype. Conclusion: Taken together, our data suggest that the in vitro system used in this study drove ATSCs toward a hepatic conversion through a subtle regulation of molecular pathways controlling stem cell properties and lineage commitment that promote mesenchymal-epithelial-transition. Adipose tissue was obtained from 3 patients undergoing partial abdominoplasty. Adipose tissue-derived stromal cells (ATSC) were isolated according to standard procedures, using the in vitro adherence property of these cells. At passage culture 4, ATSC were submitted to an in vitro hepatogenic regimen, consisting of the sequential addition of growth factors. After 1 month of in vitro differentiation, cells were harvested and their transcriptome was compared to control ATSC.
Project description:An experiment was performed in order to compare directly profiles of stromal vascular fraction of adipose tissue, testis and epidydimis, in order to rule out a possible contamination of the stromal vascular fraction of adipose tissue (SVF) by adjacent tissues (i.e. epididymis and testis). This led to the identification of a list of 2358 SVF-specific probes, which was subsequently used for further investigations. 3 dye-swap pairs, comparing 3 distinct samples of total RNA prepared from: (1) stromal vascular fraction of adipose tissue, (2) testis and (3) epidydimis.
Project description:We performed RNA-sequencing to determine the abundance of all RNA transcripts expressed by adipose-derived mesenchymal stromal cells (AMSC) under proliferating and confluent conditions.
Project description:Bone marrow (BM) has been considered so far the reference source for stromal cells (SC) originally called mesenchymal stem cells. Recently human adult adipose tissue (AT) has been reported as a valuable source for the isolation of cells exerting a mesenchymal-like phenotype. Though both BM- and AT- derived stromal cells (BMSC and ATSC) share similar immuno-phenotype and exhibit multi-lineage potential in vitro, a consensual panel of specific markers is still debated and the precise molecular mechanisms governing their differentiated fate are not fully understood. The aim of this study was to compare the genome wide expression profiles of stromal cells isolated from AT and BM and to emphasize the core of MSC stemness properties. Moreover we focused on the molecular characteristics of ATSC, attempting to reveal their specific features.We identified an overlapping dataset of 190 genes commonly regulated by ATSC and BMSC. Among these, we were able to categorize 6 key biological families that could be regarded as a stemness signature that underlie the self-renewal potential and the ability to generate progenitor cells. In particular, a pivotal role of signalling pathways along with the expression of numerous transcription regulators emerged from this study. Genes specifically modulated in ATSC, suggested that these cells posses anti-oxidative and neuroprotective properties. Taken together, these results provide new hints towards the understanding of the molecular basis of MSC maintenance and suggest that ATSC have interesting properties which could be useful for several potential clinical applications. Experiment Overall Design: Bone marrow and adipose tissue samples were respectively obtained from four long-term disease-free Hodgkin lymphoma patients, during follow-up investigation and from four patients subjected to partial abdominoplasty. In addition, we used in this study a cell line (MRC-5 cells) as control to rule out genes involved in homeostasis and emphasize genes specifically expressed in both types of mesenchymal stem cells. To have results statistically significant, we enrolled four patients for each tissue (4 chips/tissue) and we performed a technical replicate (triplicate) for the MRC-5 cells.
Project description:Adipose stromal cells are adipocytes progenitors lying in the stroma vascular fraction of adipose tissue. They can also be released into the circulation upon dietary challenges. We used microarrays to compare the global programme of gene expression between adipose stromal cells originating from murine subcutaneous and perigonadal adipose depots