[In vitro bromodeoxyuridine labeling of rat bone marrow-derived mesenchymal stem cells].
OBJECTIVE
To study the optimal dosage and timing for bromodeoxyuridine (BrdU) labeling of rat bone marrow-derived mesenchymal stem cells (MSCs) in vitro.


METHODS
Bone marrow-derived MSCs of SD rats were cultured in vitro routinely and the sixth passage was taken for identification of specific surface antigens by flow cytometry. Before reaching cell confluence, the purified MSCs were incubated with BrdU at different concentrations (5, 10, and 15 micromol/L) for different incubating time (3, 12, 24, 36, 48, and 72 h) with 10 micromol/L BrdU, to identify the optimal BrdU concentration and incubating time for cell labeling. Immunohistochemistry was performed to calculate the labeling index (LI).


RESULTS
Flow cytometry showed that MSCs expressed CD29 and CD44 but not CD11b or CD45. Incubation of the MSCs with BrdU at 10 micromol/L and for an optimal length of 48 h appeared to achieve the highest LI, both of which exceeded 98%, with the labeling identifiable in five consecutive passages.


CONCLUSIONS
The continually passaged cells are MSCs, the incubation of which with 10 micromol/L BrdU for 48 h may achieve a LI over 98% without producing obvious cell damages. The results suggest that BrdU labeling provides a feasible means for a dynamic in vivo observation of the survival, growth and differentiation of the implanted MSCs.
Feng
Shan-Wei
SW
Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
Yao
Xiao-Li
XL
Li
Zhong
Z
Liu
Tai-Yun
TY
Huang
Wen
W
Zhang
Cheng
C
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D001973:Bromodeoxyuridine; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D007202:Indicators and Reagents; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D007202:Indicators and Reagents; D001973:Bromodeoxyuridine