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Comparison of gene expression profiles between human DFAT cells and ADSCs derived from the same donors

ABSTRACT: Dedifferentiated fat (DFAT) cells, established in vitro from mature adipocytes, exhibit certain properties of multipotent mesenchymal stem/stromal cells (MSCs), such as the ability to differentiate into multiple mesenchymal lineages. Although DFAT cells exhibit properties of proliferative progeny, at present there is only limited knowledge about their MSC-specific characteristics because those cells are considered to be potential artifacts of cell culture. To elucidate the identity of DFAT cells, we compared gene expression profiles of human DFAT cells and adipose-derived stem/stromal cells (ADSCs) established using adipose tissue from the same donors. Microarray analysis showed that global mRNA expression profiles of human DFAT cells were very similar to those of ADSCs, a representative MSC, despite being committed adipocyte progenitors.

ORGANISM(S): Homo sapiens

PROVIDER: GSE47869 | GEO | 2013/06/12

SECONDARY ACCESSION(S): PRJNA208205

REPOSITORIES: GEO

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Project description:Bone tissue engineering is considered the optimal solution for the repair of large bone defects. Previous studies by our team have demonstrated the effectiveness of a "bottom-up" microtissue engineering strategy using bone marrow mesenchymal stem cells (BMSCs) to improve the utilization efficiency of seed cells. However, this strategy requires the extraction of bone marrow, which is associated with significant trauma and difficulties in obtaining the cells. Adipose-derived mesenchymal stem cells (ADSCs), obtained from aspirated adipose tissue, offer a less invasive and more accessible alternative, but their osteogenic capacity is noticeably weaker, severely limiting their application as seed cells in bone tissue engineering repair.Cell-free fat extract (CEFFE) is a cell-free liquid component extracted from adipose tissue, which exhibits beneficial properties such as promoting proliferation, antioxidation, anti-apoptosis, and angiogenesis. In this study, during the preparation of ADSC-based osteogenic microtissues, we first discovered that CEFFE significantly enhanced the osteogenic activity of ADSCs. Furthermore, we observed the formation of dense osteogenic membranes as early as day 7 of osteogenic induction. When the microtissues based on CEFFE-treated ADSCs were implanted subcutaneously in nude mice, we found a significant enhancement in their osteogenic activity. Finally, to further investigate the mechanism underlying the enhancement of ADSC osteogenic differentiation by CEFFE, we performed transcriptomic analysis of CEFFE-treated ADSCs and validated the activation of the key PI3K-Akt pathway using Western blotting.In conclusion, this study presents a novel approach for the preparation of ADSC-based osteogenic microtissues using CEFFE and provides insights into the mechanisms underlying the enhancement of osteogenic activity. These findings hold significant implications for the field of bone tissue engineering and offer potential advancements in the clinical translation of ADSC-based therapies.

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.

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Comparison of gene expression profiles between human DFAT cells and ADSCs derived from the same donors

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