MC3T3 Pre-osteoblasts were transiently transfected with plasmid DNA using LipoMag kit

Osteoclast TGF-b Receptor Signaling Induces Wnt1 Secretion and Couples Bone Resorption to Bone Formation

This article demonstrates the high efficiency of LipoMag from OZ Biosciences for DNA transfection into pre-osteoblasts

J Bone Miner Res. 2016 Jan;31(1):76-85. doi: 10.1002/jbmr.2586. Epub 2015 Aug 6.

Weivoda MM, Ruan M, Pederson L, Hachfeld C, Davey RA, Zajac J, Westendorf JJ, Khosla S, Oursler MJ.

Abstract:

Osteoblast-mediated bone formation is coupled to osteoclast-mediated bone resorption. These processes become uncoupled with age, leading to increased risk for debilitating fractures. Therefore, understanding how osteoblasts are recruited to sites of resorptionis vital to treating age-related bone loss. Osteoclasts release and activate TGF-b from the bone matrix. Here we show that osteoclastspecific inhibition of TGF-b receptor signaling in mice results in osteopenia due to reduced osteoblast numbers with no significant impact on osteoclast numbers or activity. TGF-b induced osteoclast expression of Wnt1, a protein crucial to normal bone formation,and this response was blocked by impaired TGF-b receptor signaling. Osteoclasts in aged murine bones had lower TGF-b signaling and Wnt1 expression in vivo. Ex vivo stimulation of osteoclasts derived from young or old mouse bone marrow macrophages showed no difference in TGF-b–induced Wnt1 expression. However, young osteoclasts expressed reduced Wnt1 when cultured on aged mouse bone chips compared to young mouse bone chips, consistent with decreased skeletal TGF-b availability with age.

Therefore, osteoclast responses to TGF-b are essential for coupling bone resorption to bone formation, and modulating this pathway may provide opportunities to treat age-related bone loss. © 2015 American Society for Bone and Mineral Research.

LipoMag (OZBiosciences USA, San Diego, CA, USA) was used to transfect MC3T3 preosteoblasts with a Wnt reporter plasmid containing three copies of the TCF/Lef binding site upstream of the firefly luciferase gene (TOPFlash)(31) or control DNA.

LipoMag Transfection Kit associates our most efficient lipid-based transfection reagent, DreamFect Gold with CombiMag magnetic nanoparticles, the most versatile Magnetofection™ reagent. By taking advantage of the Lipofection and Magnetofection transfection techniques, LipoMag Kit reaches even higher transfection efficiency especially in primary cells while reducing toxicity. It is suitable for all type of nucleic acids, including: plasmid DNA,linearized DNA, siRNA, oligonucleotides, double stranded RNA, mRNA, shRNA.