Quality control and monitoring for the isolation process of mesenchymal stem cells and their differentiation into osteoblasts.
We developed a method of quality control and monitoring for the isolation of mesenchymal stem cells (MSCs) from bone marrow and their differentiation into osteoblasts. After dividing the cell culture process into five groups based on cell types such as MSCs and osteoblasts, we used microarray analysis to select genes with expression profiles characteristic of each group and quantitative polymerase chain reaction for confirming the expression profiles of these genes. Comparing multiple gene expression profiles per cell from quantitative polymerase chain reaction permitted us to distinguish (1) different groups of cell culture including MSCs and osteoblasts; (2) MSCs that had differentiated cells other than osteoblasts such as chondroblasts, adipocytes, or skin-derived fibroblasts; and (3) desirable MSCs from undesirable MSCs occurring under different culture conditions. These findings suggest that it is possible to standardize MSCs and osteoblasts on the basis of multiple gene expression profiles and to check the quality of these cells. We believe that our methods can be applied to cells cultured for transplants.
Kawauchi
Satoshi
S
Mitsubishi Chemical Medience Corporation, Tokyo, Japan. Kawauchi.Satoshi@mx.medience.co.jp
Terasaki
Hiroshi
H
Katano
Masayoshi
M
Murase
Junko
J
Masuda
Yoshiko
Y
Tamura
Tomoaki
T
Shimadzu
Mitsunobu
M
D001483:Base Sequence; D018929:Cell Culture Techniques / Q000379:methods / Q000592:standards; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods* / Q000592:standards; D017931:DNA Primers / Q000235:genetics; D003663:Decision Trees; D020869:Gene Expression Profiling; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011786:Quality Control
D017931:DNA Primers