Poultry is vulnerable to bone problems throughout their
lives or
production period due to rapid growth in broilers and the active laying
cycle in layers. The calcium-sensing receptor (CaSR) is important
in calcium and bone metabolism. The objective of this study was to
investigate the effect of the CaSR ligand (l-Trp) and 1,25-dihydroxycholecalciferol
(1,25OHD3) on the regulation of proliferation and osteogenic
differentiation of chicken mesenchymal stem cells (MSCs) isolated
from the compact bones of 14-day-old Ross 308 chickens and Dekalb
pullets, which can provide cell-based evidence for the prevention
or alleviation of skeletal disorders in the poultry industry. First,
the dose- (0, 0.5, 1, 2, 5, 10, and 15 mM) and time-effects (0, 7,
and 14 days) of l-Trp on the proliferation and osteogenic
differentiation in chicken MSCs were investigated. The 5 mM l-Trp had a balanced effect between proliferation and osteogenic differentiation
in broiler and layer MSCs when differentiated for 7 days. The broiler
and layer MSCs were then treated with (1) osteogenic medium, osteogenic
medium supplemented with (2) 1 nM 1,25OHD3, (3) 2.5 mM
Ca2+, (4) 2.5 mM Ca2+ + 5 mM l-Trp
and (5) 2.5 mM Ca2+ + 5 mM l-Trp + 1 μM
NPS-2143, separately for 7 days. Results showed that the 5 mM l-Trp significantly inhibited the proliferation of broiler and
layer MSCs on day 7 (P < 0.05), but 1 nM 1,25OHD3 significantly promoted the proliferation of layer MSCs (P < 0.05). Only the 2.5 mM Ca2+ + 5 mM l-Trp group significantly increased the mineralization process
during osteogenic differentiation (P < 0.05),
and this treatment also significantly upregulated the mRNA expression
of the vitamin D receptor (VDR), β-catenin, and osteogenesis genes in broiler MSCs
(P < 0.05). The osteogenic differentiation process
in layer MSCs was faster than that in broiler MSCs. In layer MSCs,
Ca2+ alone significantly facilitated mineralization and
ALP activity after 7-day osteogenic differentiation (P < 0.05). However, the 5 mM l-Trp significantly inhibited
the differentiation and mineralization process by downregulating the
mRNA expression of CaSR, VDR, β-catenin, and
osteogenic genes (P < 0.05) in layer MSCs. Taken
together, l-Trp and 1,25OHD3 can regulate proliferation
and osteogenic differentiation in both broiler and layer MSCs depending
on the dose, treatment time, and cell proliferation and differentiation
stages.