CD271 Negative Human Dental Pulp Cells Yield Significantly More Adherent Colony Forming Cells than the Positive Phenotype

Matthew J Tomlinson

doi:10.23937/2469-570x/1410025

Abstract

Cell surface markers for isolating proliferative human dental pulp stromal cells are currently lacking. Other tissues containing mesenchymal stromal cells have been studied in greater depth and candidate markers for cell isolation identified, one such marker being CD271. Previous reports suggest CD271 as a marker for isolating dental pulp stromal cells from rat incisors. We aimed to study the utility of CD271 as a marker for isolating human dental pulp stromal cells. CD271 positive cells from both third molar dental pulp and bone marrow mononuclear cells were isolated by magnetic separation followed by in vitro expansion. Phenotypic analysis was performed by flow cytometry. Our data showed that although CD271 is present in the adult molar dental pulp, significantly greater numbers of colonies are produced from the CD271 negative fraction. The opposite was seen with bone marrow mononuclear cells; all colony forming cells were derived from the CD271 positive fraction. Phenotypic analysis of expanded CD271 negative cells showed that these cells are identical to dental pulp stromal cells isolated by non-selective plastic adherence. We conclude that, in contrast to bone marrow, CD271 is not a positive selection marker for the predominant colony forming cell types from human molar dental pulp.

Highlights

Human dental pulp stromal cells are plastic adherent cells capable of in vitro colony formation, mineralisation and demonstrable mesenchymal differentiation potential [1,2]
Analysis of cultured Human dental pulp stromal cells (hDPSCs), derived from plastic adherence, revealed that CD271 expression decreased to very low levels upon induction into 2 dimensional culture with an average expression of 0.2% across passages 2 to 4 (Figure 1b)
Cells from whole digested dental pulp (DP) were sorted for expression of CD271, isolated cells were used in colony formation unit fibroblast (CFU-F) assays, CD271+ and CD271- cells were plated at the ratio of cells found within the DP

Summary

Introduction

Human dental pulp stromal cells (hDPSCs) are plastic adherent cells capable of in vitro colony formation, mineralisation and demonstrable mesenchymal differentiation potential [1,2]. They are of interest for tissue engineering and regenerative medicine applications, in particular dental and orthopaedic procedures, due to the potential for banking cells from the deciduous dentition [2,3]. Within the dental pulp (DP), hDPSCs are present in comparatively low numbers, but undergo expansion in vitro and are phenotypically similar to mesenchymal stromal cells (MSCs) from other tissues [4,5]. One interesting marker is the neutrophin receptor CD271 which has shown specificity for colony forming cells from BM [11,12,13]

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