The floating yellowish lipid and remnants were discarded and the infranatant
was transferred into a sterile 15 ml centrifuge tube and centrifuged at 1200 rpm for 10 min.
The supernatant and infranatant
were discarded, and the bottom 2 ml of fat left in the syringes was transferred to 1-ml syringes.
The floating adipocytes were collected and washed and the infranatant
was removed and centrifuged at 500 x g, for 5 minutes to pellet the stromal vascular fraction (SVF).
separated by funnel after adding ethyl ether (20 mL; Sigma-Aldrich, Saint Louis, MO, USA) was evaporated.
was removed by gentle aspiration using a plastic Pasteur pipette.
The collected infranatant
fraction is then brought back to its original volume, and the cholesterol content is then measured by the Abell-Kendall method.
* If the specimen is overtly lipemic (lipemic samples can also be ultracentrifuged prior to analysis by indirect measurement of the aqueous infranatant
) or hyperviscous.
At the end of the incubation period, an aliquot of the infranatant
was removed for enzymatic determination of glycerol released into the incubation medium (KATAL, Belo Horizonte, MG, Brazil).
fraction can now be reconstituted to the original volume with 0.15 M NaCl after the supernatant VLDL- and chylomicron-containing fractions have been quantitatively removed.
Therefore, the analysis of the clear infranatant
solution after ultracentrifugation will not give accurate triglyceride results.
We isolated the fraction of density > 1.006 g/mL (infranatant
) and measured TC and HDL-C as described above.
containing immature adipocytes was discarded and then the remained mature adipocytes were collected.