10.6084/m9.figshare.3160450.v2
Amr Alraies
Amr
Alraies
Figure 3. Osteogenic, chondrogenic and adipogenic differentiation of DPPCs.
figshare
2016
Stem Cell Differentiation
Cell Biology
2016-04-14 08:19:11
Figure
https://figshare.com/articles/figure/Figure_3/3160450
<br>
<p>Osteogenic,
chondrogenic and adipogenic differentiation of DPPCs. Tri-lineage differentiation analysis was
compared between high proliferative capacity, A3 (30 PDs) and low proliferative
capacity DPPCs, A1 (11 PDs) and B1 (15 PDs).
(A) Osteogenesis was confirmed for each DPPC population by the detection
of alizarin red staining for mineralised calcium nodules. No staining was observed in control DPPC
cultures in the absence of osteogenic medium.
RT-PCR analysis of osteogenic markers (right panel) indicated increased osteocalcin
(OCN) and osteopontin (OPN) expression for DPPCs cultured in osteogenic media. Cells maintained in both osteogenic and
control media expressed Runx2. (B)
Chondrogenesis was only particularly evident with high proliferative capacity,
A3, which exhibited positive Safranin O staining for high proteoglycan
content. No Safranin O staining was
observed with the low proliferative capacity DPPCs, A1 and B1. (C) Adipogenesis was only particularly
evident with high proliferative capacity, A3, which exhibited positive Oil Red
O staining for intracellular lipid-rich vacuole accumulation. No
Oil Red O staining was observed with the low proliferative capacity DPPCs, A1
and B1; or in control cultures in the absence of adipogenic medium. RT-PCR analysis of adipogenic markers (right
panel) indicated increased expression of the early adipogenic marker, PPARγ, in
all DPPCs maintained in both adipogenic and control cultures; whilst the late
adipogenic marker, lipoprotein lipase (LPL), was only expressed in high
proliferative capacity, A3, in adipogenic media. Scale bars = 100 µm. β-actin served as the reference housekeeping
gene.</p>