Isolation and Characterization of Buccal Fat Pad and Dental Pulp MSCs from the Same Donor.

Tullio Genova,Fabio Tasinato,Ilaria Roato,Sara Petrillo,Luca Mela,Davide Cavagnetto,Luca Munaron,Massimo Carossa,Federico Mussano,Federico Alessandro Ruffinatti

doi:10.3390/biomedicines9030265

Tullio Genova, Fabio Tasinato + Show 8 more

Open Access

https://doi.org/10.3390/biomedicines9030265

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Abstract

Mesenchymal stem cells (MSCs) can be harvested from different sites in the oral cavity, representing a reservoir of cells useful for regenerative purposes. As direct comparisons between at least two types of MSCs deriving from the same patient are surprisingly rare in scientific literature, we isolated and investigated the osteoinductive potential of dental pulp stem cells (DPSCs) and buccal fat pad stem cells (BFPSCs). MSCs were isolated from the third molar dental pulp and buccal fat pads of 12 patients. The number of viable cells was quantified through manual count. Proliferation and osteodifferentiation assays, flow cytometry analysis of cell phenotypes, and osteocalcin release in vitro were performed. The isolation of BFPSCs and DPSCs was successful in 7 out of 12 (58%) and 3 out of 12 (25%) of retrieved samples, respectively. The yield of cells expressing typical stem cell markers and the level of proliferation were higher in BFPSCs than in DPSCs. Both BFP-SCs and DPSCs differentiated into osteoblast-like cells and were able to release a mineralized matrix. The release of osteocalcin, albeit greater for BFPSCs, did not show any significant difference between BFPSCs and DPSCs. The yield of MSCs depends on their site of origin as well as on the protocol adopted for their isolation. Our data show that BFP is a valuable source for the derivation of MSCs that can be used for regenerative treatments.

Highlights

Human mesenchymal stem cells (MSCs) are the fundament of any tissue engineering approach aiming at regenerating mineralized tissues [1]
Success Rates in Isolating MSCs from buccal fat pad (BFP) and dental pulp (DP). Both the buccal fat pad stem cells (BFPSCs) and dental pulp stem cells (DPSCs) adhered to the cell culture dishes, and their isolation was evident from an optical microscope observation of cell adhesion and formation of colonies
DP As reported in Figure 1A, the success rate in isolating stem cells was higher for BFP than for DP. Both the BFPSCs and DPSCs adhered to the cell culture dishes, The growth of BFPSCs and DPSCs was assessed at the first passage, showing a sigand their isolation was evident from an optical microscope observation of cell adhesion nificantly reduced number Cell of viable cells was in cultures of DP compared to BFP, as cells shown in and formation of colonies

Summary

Introduction

Human mesenchymal stem cells (MSCs) are the fundament of any tissue engineering approach aiming at regenerating mineralized tissues [1]. One of the first sites to derive MSCs has been bone marrow, but extraction is hindered by difficult accessibility and a frequently painful procedure [2]. Alternative sources of MSCs have been discovered, such as adipose tissue and dental pulp (DP), which can be and conspicuously harvested [3]. Adipose-derived stem cells (ASCs) are a plastic-adherent [4], possess a multipotent cell population attainable during liposuction, and rely on an impressive corpus of supporting research published since 2001 [5] reporting on their capability to differentiate into different tissues, such as bone and cartilage [6,7,8]. Extracted teeth have become an interesting source of MSCs since the first

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