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MSC to adipocyte (human): Difference between revisions

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|timepoint_design=Early focus
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|tissue_cell_type=Mesenchymal>>adipose
|tissue_cell_type=Mesenchymal>>adipose
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Revision as of 15:57, 12 February 2015

Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:FILE
Link to TET:TET
Sample providers :Peter Arner
Germ layer:mesoderm
Primary cells or cell line:primary cells
Time span:14 days
Number of time points:15


Overview

Adipose tissue can account for between 5% (lean athletes) and 60% (morbidly obese) of total body mass, making it one of the most plastic organs in the body [8]. In response to changed nutritional status, both adipocyte cell number and size change and even under stable conditions as much as 10% of the adipocytes are turned over annually [1,2]. Thus the birth of new adipocytes from precursor cells (adipogenesis) is central for a functional fat tissue.

References:
[1] Dynamics of fat cell turnover in humans. Spalding KL et al, Nature, 2008, vol 453, p783-787. PMID:18454136
[2] Adipocyte turnover:relevance to human adiopose tissue morphology- Arner E et al, Diabetes, 2010,59(1):105-9. PMID:19846802
[8] Cawthorn WP, Scheller EL, MacDougald OA., Adipose tissue stem cells meet preadipocyte commitment: going back to the future. J Lipid Res. 2012 Feb;53(2):227-46.

Sample description

We are using two different models of human adipogenesis in the FANTOM5 study. Both originate from the stromal-vascular fraction (SVF) of human subcutaneous adipose tissue. The first model, here called preadipocytes, consists of cells differentiated in vitro directly after SVF isolation from adipose tissue. This means only later stages of differentiation can be monitored as samples in the beginning contain other cell types, like immune and endothelial cells, that contaminates the results. These contaminants die off after approximately 3-4 days. For this model we have taken tissue from four donors and sampled differentiation at time-points 4, 8 and 12 days after differentiation start. We have also isolated mature adipocytes from the adipose tissue of these donors. For the other model system, here called human adipose-derived mesenchymal stem cells (hASC), we have propagated cells from the SVF in vitro thus selecting for a stem cell/progenitor population. These cells can be expanded and cultured for several passages. This gives us cells with a homogenous genetic background and since contaminants are removed from the beginning, differentiation can be monitored from start to finish. Here we have collected triplicate samples at 0, 15, 30, 45 min, 1 h, 1 h 20 min, 1 h 40 min, 2 h, 2 h 30 min, 3 h, 12 h and 1, 2, 4, 8, 12 and 14 days after differentiation start.



Figure 1. Sample collection scheme for the different model system. Sample collection time points are indicated as well as the removal of some components of the differentiation cocktail. IBMX=3-isobutyl-1-methylxanthine, Dexam.=dexamethasone, Rosig.=Rosiglitazone.

Quality control

A key aspect of adipocyte maturation is the accumulation of lipid in intracellular lipid droplets. The accumulation of lipids can be seen as morphological changes in the microscope at low magnification (10x) and can be further visualized using lipid stains such as bodipy (shown below for day 8 and 12).


Figure 2: Lipid accumulation during differentiation. hASC cells were stained with Hoechst (nuclei, blue) day 0, 8 and 12 and Bodipy (neutral lipids, green) at day 8 and 12 during differentiation. An overlay of phase contrast and fluorescent image is shown.

Marker gene expression:

Preadipocyte, CAGE:



hASC, CAGE:


hASC, qPCR:



Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples


13229-141I7mesenchymal stem cells (adipose derived), adipogenic induction00hr00minbiol_rep1
13230-141I8mesenchymal stem cells (adipose derived), adipogenic induction00hr00minbiol_rep2
13231-141I9mesenchymal stem cells (adipose derived), adipogenic induction00hr00minbiol_rep3
13232-142A1mesenchymal stem cells (adipose derived), adipogenic induction00hr15minbiol_rep1
13233-142A2mesenchymal stem cells (adipose derived), adipogenic induction00hr15minbiol_rep2
13234-142A3mesenchymal stem cells (adipose derived), adipogenic induction00hr15minbiol_rep3
13235-142A4mesenchymal stem cells (adipose derived), adipogenic induction00hr30minbiol_rep1
13236-142A5mesenchymal stem cells (adipose derived), adipogenic induction00hr30minbiol_rep2
13238-142A7mesenchymal stem cells (adipose derived), adipogenic induction00hr45minbiol_rep1
13239-142A8mesenchymal stem cells (adipose derived), adipogenic induction00hr45minbiol_rep2
13240-142A9mesenchymal stem cells (adipose derived), adipogenic induction00hr45minbiol_rep3
13241-142B1mesenchymal stem cells (adipose derived), adipogenic induction01hr00minbiol_rep1
13242-142B2mesenchymal stem cells (adipose derived), adipogenic induction01hr00minbiol_rep2
13243-142B3mesenchymal stem cells (adipose derived), adipogenic induction01hr00minbiol_rep3
13247-142B7mesenchymal stem cells (adipose derived), adipogenic induction01hr40minbiol_rep1
13248-142B8mesenchymal stem cells (adipose derived), adipogenic induction01hr40minbiol_rep2
13249-142B9mesenchymal stem cells (adipose derived), adipogenic induction01hr40minbiol_rep3
13250-142C1mesenchymal stem cells (adipose derived), adipogenic induction02hr00minbiol_rep1
13251-142C2mesenchymal stem cells (adipose derived), adipogenic induction02hr00minbiol_rep2
13253-142C4mesenchymal stem cells (adipose derived), adipogenic induction02hr30minbiol_rep1
13255-142C6mesenchymal stem cells (adipose derived), adipogenic induction02hr30minbiol_rep3
13256-142C7mesenchymal stem cells (adipose derived), adipogenic induction03hr00minbiol_rep1
13258-142C9mesenchymal stem cells (adipose derived), adipogenic induction03hr00minbiol_rep3
13263-142D5mesenchymal stem cells (adipose derived), adipogenic inductionday01biol_rep2
13264-142D6mesenchymal stem cells (adipose derived), adipogenic inductionday01biol_rep3
13265-142D7mesenchymal stem cells (adipose derived), adipogenic inductionday02biol_rep1
13266-142D8mesenchymal stem cells (adipose derived), adipogenic inductionday02biol_rep2
13267-142D9mesenchymal stem cells (adipose derived), adipogenic inductionday02biol_rep3
13268-142E1mesenchymal stem cells (adipose derived), adipogenic inductionday04biol_rep1
13270-142E3mesenchymal stem cells (adipose derived), adipogenic inductionday04biol_rep3
13271-142E4mesenchymal stem cells (adipose derived), adipogenic inductionday08biol_rep1
13272-142E5mesenchymal stem cells (adipose derived), adipogenic inductionday08biol_rep2
13273-142E6mesenchymal stem cells (adipose derived), adipogenic inductionday08biol_rep3
13274-142E7mesenchymal stem cells (adipose derived), adipogenic inductionday12biol_rep1
13275-142E8mesenchymal stem cells (adipose derived), adipogenic inductionday12biol_rep2
13276-142E9mesenchymal stem cells (adipose derived), adipogenic inductionday12biol_rep3
13277-142F1mesenchymal stem cells (adipose derived), adipogenic inductionday14biol_rep1
13279-142F3mesenchymal stem cells (adipose derived), adipogenic inductionday14biol_rep3
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