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.
Huang
W
W
Plastic Surgery Section, VA Greater Los Angeles Healthcare System, Los Angeles, California, 90073, USA.
Carlsen
B
B
Rudkin
G H
GH
Shah
N
N
Chung
C
C
Ishida
K
K
Yamaguchi
D T
DT
Miller
T A
TA
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