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

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

International journal of hematology (2008-03-05)

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

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.

MATERIALS

Product Number

Brand

Product Description

M0446

Sigma-Aldrich

StableCell^™ MEM, With Earle′s salts, stable glutamine, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture

M6199

Sigma-Aldrich

StableCell^™ MEM, Alpha Modification, With stable glutamine and sodium bicarbonate, without ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture

M1018

Sigma-Aldrich

Minimum Essential Medium Eagle, With Hanks′ salts, L-glutamine and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture

F0392

Sigma-Aldrich

Fetal Bovine Serum, USA origin, Dialyzed by ultrafiltration against 0.15 M NaCl, sterile-filtered, suitable for cell culture

F3297

SAFC

Fetal Bovine Serum, Australia origin, γ-irradiated, suitable for cell culture, suitable for hybridoma

M4642

Sigma-Aldrich

Minimum Essential Medium Eagle, With Hanks′ salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture

M0769

SAFC

Minimum Essential Medium Eagle, AutoMod^™, with Earle′s salts, without L-glutamine and sodium bicarbonate, powder, suitable for cell culture

M4655

Sigma-Aldrich

Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture

M0268

Sigma-Aldrich

Minimum Essential Medium Eagle, With Earle′s salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture

F4135

Sigma-Aldrich

Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture, suitable for insect cell culture, suitable for hybridoma

M0894

Sigma-Aldrich

Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine and sodium pyruvate, without ribonucleosides, deoxyribonucleosides and sodium bicarbonate, powder, suitable for cell culture

F6765

Sigma-Aldrich

Fetal Bovine Serum, USA origin, Charcoal Stripped, sterile-filtered, suitable for cell culture

M0643

Sigma-Aldrich

Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine, and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture

F2442

Sigma-Aldrich

Fetal Bovine Serum, USA origin, sterile-filtered, suitable for cell culture, suitable for hybridoma

M2645

SAFC

Minimum Essential Medium Eagle, HEPES Modification, with Earle′s salts, L-glutamine and 25 mM HEPES, without sodium bicarbonate, powder, suitable for cell culture

M0518

Sigma-Aldrich

Minimum Essential Medium Eagle, Joklik Modification, with L-glutamine, without calcium chloride and sodium bicarbonate, suitable for cell culture

M0644

Sigma-Aldrich

Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine, ribonucleosides and deoxyribonucleosides, without sodium bicarbonate, powder, suitable for cell culture

F7524

Sigma-Aldrich

Fetal Bovine Serum, non-USA origin, sterile-filtered, suitable for cell culture

F9665

Sigma-Aldrich

Fetal Bovine Serum, Heat Inactivated, non-USA origin, sterile-filtered, suitable for cell culture

F9423

Sigma-Aldrich

Fetal Bovine Serum, Australia origin, suitable for, USDA approved, sterile-filtered, suitable for cell culture, suitable for hybridoma

12107C

Sigma-Aldrich

Fetal Bovine Serum, USA origin, γ-irradiated by SER-TAIN process, sterile-filtered, suitable for cell culture

12106C

Sigma-Aldrich

Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture

12103C

Sigma-Aldrich

Fetal Bovine Serum, USA origin, suitable for cell culture

12306C

Sigma-Aldrich

Fetal Bovine Serum, non-USA origin, from USDA approved countries, Heat Inactivated, sterile-filtered, suitable for cell culture

12003C

Sigma-Aldrich

Fetal Bovine Serum, Australia origin, USDA approved, sterile-filtered, suitable for cell culture

12007C

Sigma-Aldrich

Fetal Bovine Serum, Australia origin, γ-irradiated, sterile-filtered, suitable for cell culture

M3024

Sigma-Aldrich

Minimum Essential Medium Eagle, with Earle′s salts and non-essential amino acids, without L-glutamine, phenol red and sodium bicarbonate, Modified, powder, suitable for cell culture

12006C

Sigma-Aldrich

Fetal Bovine Serum, Australia origin, USDA approved, Heat Inactivated, suitable for cell culture

M2414

Sigma-Aldrich

Minimum Essential Medium Eagle, Modified, with Earle′s salts and reduced NaHCO₃. without L-glutamine, liquid, sterile-filtered, suitable for cell culture

M0275

Sigma-Aldrich

Minimum Essential Medium Eagle, 10 ×, With Earle′s salts, without L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture