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. 2008 Apr;87(3):239-45.
doi: 10.1007/s12185-008-0048-9. Epub 2008 Mar 4.

Identification of genes potentially involved in supporting hematopoietic stem cell activity of stromal cell line MC3T3-G2/PA6

Natsumi Shimizu  1 , Shinichi Noda, Kazufumi Katayama, Hitoshi Ichikawa, Hiroaki Kodama, Hiroyuki Miyoshi
Affiliations

Identification of genes potentially involved in supporting hematopoietic stem cell activity of stromal cell line MC3T3-G2/PA6

Natsumi Shimizu et al. Int J Hematol. 2008 Apr.

Abstract

Although coculture of hematopoietic stem cells (HSCs) with stromal cells is a useful system to study hematopoiesis in the niche, little is known regarding the precise cellular and molecular mechanisms of maintaining HSCs through cell-cell interactions. The murine preadipose stromal cell line MC3T3-G2/PA6 (PA6) has been demonstrated to support HSCs in vitro. In this study, microarray analysis was performed on PA6 cells and HSC-nonsupporting PA6 subclone cells to identify genes responsible for supporting HSC activity. Comparison of gene expression profiles revealed that only 144 genes were down-regulated by more than twofold in PA6 subclone cells. Of these down-regulated genes, we selected 11 candidate genes and evaluated for the maintenance of HSC function by overexpressing these genes in PA6 subclone cells. One unknown gene, 1110007F12Rik (also named as Tmem140), which is predicted to encode an integral membrane protein, demonstrated a partial restoration of the defect in HSC-supporting activity.

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Figures

Fig. 1
Fig. 1
Effect of over expression of candidate genes on the CFC frequency. A total of 80 CD34−KSL HSCs (10 cells/well) were cocultured with PA6 cells, PA6 S-2 cells, or PA6 S-2 cells expressing the indicated genes for 10 days and then subjected to the CFC assay. Individual colonies were scored according to their morphology. Data represent the average of two independent experiments
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