Effect of serial passage on gene expression in MC3T3-E1 preosteoblastic cells: a microarray study.
The osteoblastic function of mouse preosteoblastic MC3T3-E1 cells, as measured by alkaline phosphatase activity and osteocalcin secretion, decreases after serial passage. To uncover genes responsible for decreased osteoblastic function in high-passage cells, we have studied passage-dependent change of gene expression in MC3T3-E1 cells. Changes in the expression pattern of 2000 selected genes were examined simultaneously by comparing mRNA levels between MC3T3-E1 cells at passage 20 and passage 60 using the cDNA microarray analysis. Significant changes in the steady-state abundance of 27 mRNAs were observed in response to different passage numbers, including 17 known genes, 4 ESTs with homology to known genes, and 6 genes with no previously described function or homology. Northern blot analysis was used to verify and quantify the expression of selected genes, and revealed a significant higher level of up- and down-regulation compared to microarray data. These results indicate the existence of a significant change in gene expression in osteoblastic cells undergoing serial passages. Such changes might be responsible for a reduction in bone regeneration in older osteoblasts. Potential roles of selected genes in bone aging are discussed.
Plastic Surgery Section, VA Greater Los Angeles Healthcare System, Los Angeles, California, 90073, USA.
W
W
Huang
B
B
Carlsen
G H
GH
Rudkin
N
N
Shah
C
C
Chung
K
K
Ishida
D T
DT
Yamaguchi
T A
TA
Miller
D000375:Aging; D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D020869:Gene Expression Profiling; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D013234:Stem Cells / Q000378:metabolism; D014158:Transcription, Genetic
D012333:RNA, Messenger