Recently, gingival margin-derived stem/progenitor cells isolated via
STRO-1/magnetic activated cell sorting (MACS) showed remarkable periodontal
regenerative potential in vivo. As a second-stage investigation, the present
study's aim was to perform in vitro characterisation and comparison of the
stem/progenitor cell characteristics of sorted STRO-1-positive (MACS+) and
STRO-1-negative (MACS−) cell populations from the human free gingival margin.
Cells were isolated from the free gingiva using a minimally invasive technique
and were magnetically sorted using anti-STRO-1 antibodies. Subsequently, the
MACS+ and MACS− cell fractions were characterized by flow cytometry for
expression of CD14, CD34, CD45, CD73, CD90, CD105, CD146/MUC18 and STRO-1.
Colony-forming unit (CFU) and multilineage differentiation potential were
assayed for both cell fractions. Mineralisation marker expression was examined
using real-time polymerase chain reaction (PCR). MACS+ and MACS− cell
fractions showed plastic adherence. MACS+ cells, in contrast to MACS− cells,
showed all of the predefined mesenchymal stem/progenitor cell characteristics
and a significantly higher number of CFUs (P<0.01). More than 95% of MACS+
cells expressed CD105, CD90 and CD73; lacked the haematopoietic markers CD45,
CD34 and CD14, and expressed STRO-1 and CD146/MUC18. MACS− cells showed a
different surface marker expression profile, with almost no expression of CD14
or STRO-1, and more than 95% of these cells expressed CD73, CD90 and
CD146/MUC18, as well as the haematopoietic markers CD34 and CD45 and CD105.
MACS+ cells could be differentiated along osteoblastic, adipocytic and
chondroblastic lineages. In contrast, MACS− cells demonstrated slight
osteogenic potential. Unstimulated MACS+ cells showed significantly higher
expression of collagen I (P<0.05) and collagen III (P<0.01), whereas MACS−
cells demonstrated higher expression of osteonectin (P<0.05; Mann–Whitney).
The present study is the first to compare gingival MACS+ and MACS− cell
populations demonstrating that MACS+ cells, in contrast to MACS− cells,
harbour stem/progenitor cell characteristics. This study also validates the
effectiveness of the STRO-1/MACS+ technique for the isolation of gingival
stem/progenitor cells. Human free gingival margin-derived STRO-1/MACS+ cells
are a unique renewable source of multipotent stem/progenitor cells