A Novel Gene (drad‐1) Expressed in Hematopoiesis‐Supporting Stromal Cell Lines, ST2, PA6 and A54 Preadipocytes: Use of mRNA Differential Display

Abstract

We established a differentiation-inducible preadipocyte cell line, designated A54 preadipocytes, from C3H10T1/2 (10T1/2) mouse embryo fibroblasts. A54 preadipocytes had marked hematopoiesis-supporting ability in vitro but this ability was lost after terminal differentiation to adipocytes. In this study, to identify molecules that contribute to the hematopoiesis-supporting ability of A54 preadipocytes, we screened genes that were differentially expressed in A54 preadipocytes and isolated seven novel genes by reverse transcriptase polymerase chain reaction mRNA differential display. An RNase protection assay confirmed that one of these genes was expressed at high levels in parent 10T1/2 cells and A54 preadipocytes but to a much lesser extent in fully differentiated A54 adipocytes. This gene was defined as a gene that was downregulated during adipocyte differentiation-1 (drad-1). The size of drad-1 mRNA was 8.2 kb, and the gene was expressed in other mouse preadipocytes, namely, ST2 and PA6 cells, that have hematopoiesis-supporting ability. Moreover, drad-1 was also found to be expressed in bone marrow in vivo. The function of the protein encoded by drad-1 is unknown, but the expression of the gene may be useful as a molecular marker of adipocyte differentiation.

Cell and gene therapy using mesenchymal stem cells (MSCs)

“…Previous studies have shown that preadipocytes have a higher ability to support hematopoiesis than other kinds of stromal cell components in vitro [12,13]. Our results of gene expression profile revealed up-regulation of critical cytokines for hematopoiesis such as SCF and SDF-1 in preadipocyte A54 cells.…”

“…In preadipocyte A54 cells, 201 genes were up-regulated, including genes known to be involved in adipocyte differentiation such as genes encoding C/EBPa, C/EBPd, PPAR-g, PAI-I, and Frizzled-1 [10]. Myoblasts M1601 cells showed 137 upregulated genes, including ones related to skeletal muscle differentiation such as genes encoding MyoD, MLC1F, askeletal actin, myosin heavy chain, and myosin light chain [11] as well as genes related to cardiac muscle differentiation such as genes encoding a-cardiac actin, cardiac troponin C, and troponin T2 [12].…”

Human Hematopoietic Stem Cells Can Survive In Vitro for Several Months

“…We used the mouse-derived cell lines, OP9 and C3H10T1/2, as feeder cells. Both OP9 [10–15] and C3H10T1/2 [10, 16–18] have been used in many studies to maintain in vitro hematopoiesis. As we show below, both OP9 and C3H10T1/2 cells supported long-lasting in vitro hematopoiesis from human hematopoietic stem cells.…”