Journal of Oral Science
Online ISSN : 1880-4926
Print ISSN : 1343-4934
ISSN-L : 1343-4934
Effect of collagenase concentration on the isolation of small adipocytes from human buccal fat pad
Niina TsurumachiDaisuke AkitaKoichiro KanoTaro MatsumotoTaku ToriumiTomohiko KazamaYoshinao OkiYoko Saito-TamuraMorio TonogiNoriyoshi ShimizuMasaki Honda
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JOURNALS FREE ACCESS Advance online publication

Article ID: 16-0786

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Abstract

Dedifferentiated fat (DFAT) cells were isolated from mature adipocytes using the ceiling culture method. Recently, we successfully isolated DFAT cells from adipocytes with a relatively small size (<40 μm). DFAT cells have a higher osteogenic potential than that of medium adipocytes. Therefore, the objective of this study was to determine the optimal concentration of collagenase solution for isolating small adipocytes from human buccal fat pads (BFPs). Four concentrations of collagenase solution (0.01%, 0.02%, 0.1%, and 0.5%) were used, and their effectiveness was assessed by the number of small adipocytes and DFAT cells isolated. The total number of floating adipocytes that dissociated with 0.02% collagenase was 2.5 times of that dissociated with 0.1% collagenase. The number of floating adipocytes with a diameter of ≤29 μm that dissociated with 0.02% collagenase was thrice of those dissociated with 0.1% and 0.5% collagenase. The number of DFAT cells that dissociated with 0.02% collagenase was 1.5 times of that dissociated with 0.1% collagenase. In addition, DFAT cells that dissociated with 0.02% collagenase had a higher osteogenic differentiation potential than those that dissociated with 0.1% collagenase. These results suggest that 0.02% is the optimal collagenase concentration for isolating small adipocytes from BFPs.

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