Small Buccal Fat Pad Cells Have High Osteogenic Differentiation Potential

Abstract

Dedifferentiated fat (DFAT) cells derived from mature adipocytes have mesenchymal stem cells' (MSCs) characteristics. Generally, mature adipocytes are 60-110 μm in diameter; however, association between adipocyte size and dedifferentiation efficiency is still unknown. This study, therefore, investigated the dedifferentiation efficiency of adipocytes based on cell diameter. Buccal fat pad was harvested from five human donors and dissociated by collagenase digestion. After exclusion of unwanted stromal cells by centrifugation, floating adipocytes were collected and their size distribution was analyzed. The floating adipocytes were then separated into two groups depending on cell size using 40- and 100-μm nylon mesh filters: cell diameters less than 40 μm (small adipocytes: S-adipocytes) and cell diameters of 40-100 μm (large adipocytes: L-adipocytes). Finally, we evaluated the efficiency of adipocyte dedifferentiation and then characterized the resultant DFAT cells. The S-adipocytes showed a higher capacity to dedifferentiate into DFAT cells (S-DFAT cells) compared to the L-adipocytes (L-DFAT cells). The S-DFAT cells also showed a relatively higher proportion of CD146-positive cells than L-DFAT cells, and exhibited more osteogenic differentiation ability based on the alkaline phosphatase activity and amount of calcium deposition. These results suggested that the S- and L-DFAT cells had distinct characteristics, and that the higher dedifferentiation potential of S-adipocytes compared to L-adipocytes gives the former group an advantage in yielding DFAT cells.

Buccal Fat Pad as a Potential Source of Stem Cells for Bone Regeneration: A Literature Review

“…All studies derived stem cells from human volunteers except for a study by Niada et al, where BFPSCs were derived from swine and compared to SC-AdSCs [13]. Three of the 10 studies focused on BFP-DFAT cells [14–16]. One experiment focused on the size of DFAT cells and compared small cells (<40  μ m) with large cells (40–100  μ m) based on the characteristics for MSCs [16].…”

“…Three of the 10 studies focused on BFP-DFAT cells [14–16]. One experiment focused on the size of DFAT cells and compared small cells (<40  μ m) with large cells (40–100  μ m) based on the characteristics for MSCs [16]. …”

“…Unlike terminally differentiated adipocytes, DFAT cells have significant and steady proliferation capability [30–33]. In contrast to AdSCs, DFAT cells have been shown to have a more homogeneous cell population [15, 16, 30]. In addition, a greater number of DFAT cells can be produced from a given amount of fat tissue [34, 35].…”

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

“…Cell phenotypes were identified using flow cytometry as described previously (47,60). Cells were suspended at a density of 1 × 10 6 cells per tube and then incubated with antibodies against the following antigens: cluster of differentiation (CD) 44-hyaluronate receptor, phagocytic glycoprotein-1-(CD44-FITC; BD Biosciences, San Jose, CA, USA), CD73/Ecto-5-prime-nucleotidase (NT5E) (CD73-PE; Biolegend, San Diego, CA, USA), CD90/Thy-1 (CD90-APC; BD Biosciences), CD105/endoglin (CD105-FITC; Biolegend), CD146 -the cell surface glycoprotein MUC18, the melanoma cell adhesion molecule (MCAM)-(CD146-PE; BD Biosciences), the neurotrophin receptor P75 (P75) -CD271, nerve growth factor receptor (NGFR) -(P75-APC; Biolegend), stage-specific embryonic antigen (SSEA)-4 (SSEA-4-Alexa Fluor 647; BD Biosciences), and human natural killer (HNK)-1-the neural cell adhesion molecule (NCAM), CD57-(Sigma-Aldrich).…”

“…Migrated cells were plated at 1 × 10 5 cells per well in 6-well plates and cultured for 1, 3, 5, or 7 days in neural induction medium or mesenchymal stem cell (MSC) growth medium consisting of the alpha modification of Eagle's medium (α-MEM; Wako) supplemented with 10% fetal bovine serum, 50 U/mL penicillin, and 50 μg/mL streptomycin (Wako). Cell proliferation was determined by water-soluble tetrazolium salt (WST)-8 assay using a Cell Counting Kit-8 (CCK-8; Dojindo) according to the manufacturer's instructions as described previously (47,60).…”