ABSTRACT
Objective To study the osteogenic effects of induced endothelial cell(EC)on bone marrow stromal cells(BMSCs)of rabbits in co-culture condition.Methods BMSCs were obtained from rabbits by density gradient centrifugation.The adhesive ceils were preserved to passage in culture.The cultured cells were divided into four groups:group A(BMSCs),group B(BMSCs osteogenic induction),group C(EC induction)and group D (co-culture of induced BMSCs and EC).The cell morphology,immunofluorescence,cell proliferation,alkaline pbosphatase(ALP)activity,osteocalcin synthesis were observed to determine the effects of induced autologous EC on the osteogenic potential and cellular compatibility of BMSCs.Results The immunochemical staining showed that the BMSCs were induced into EC in group C.The cellular compatibility of BMSCs and EC was good.The ALP activity and osteocalcin content were obviously higher in group D than in any other groups(P＜0.05).The cell proliferation difference was not obvious between groups(P＞0.05).Conclusions The cellular compatibility of induced osteoblasts and induced EC is perfect.The ECs can significantly increase the viability and ALP activity of induced osteoblasts.
ABSTRACT
Effects of oxidized low-density lipoprotein (ox-LDL), l-alpha-stearoyl-lysophosphatidylcholine (LPC), on intracellular Ca2+ concentration were examined in mouse endothelial cells by measuring intracellular Ca2+ concentration ([Ca2+]i) with fura 2-AM and reverse transcription-polymerase chain reaction (RT-PCR). LPC increased [Ca2+]i under the condition of 1.5 mM [Ca2+]o but did not show any effect under the nominally Ca2+-free condition. Even after the store depletion with 30microM 2,5-di-tert- butylhydroquinone (BHQ) or 30microM ATP, LPC could still increase the [Ca2+]i under the condition of 1.5 mM [Ca2+]o. The time required to increase [Ca2+]i (about 1 minute) was longer than that for ATP-induced [Ca2+]i increase (10-30 seconds). LPC-induced [Ca2+]i increase was completely blocked by 1microM La3+. Transient receptor potential channel(trpc) 4 mRNA was detected with RT-PCR. From these results, we suggest that LPC increased [Ca2+]i via the increase of Ca2+ influx through the Ca2+ routes which exist in the plasma membrane.
Subject(s)
Animals , Mice , Adenosine Triphosphate , Cell Membrane , Endothelial Cells , Lipoproteins , Lysophosphatidylcholines , RNA, MessengerABSTRACT
In order to determine the precise mechanism of the interactions between different types of cells, which are common phenomena in tissues and organs, the importance of coculture techniques are becoming increasingly important. In the area of cardiology, artificial arteries have been developed, based on the understanding of physiological communication of the arterial smooth muscle cells (SMC), endothelial cells (EC), and the extracellular matrix (ECM). In the study of atherosclerosis, the modification of low-density lipoprotein (LDL), which result in the recruitment and accumulation of white blood cells, especially, monocytes/macrophages, and foam cell formation, are hypothesized. Although there are well known animal models, an in vitro model of atherogenesis with a precisely known atherogenesis mechanism has not yet been developed. In this paper, an arterial wall reconstruction model using rabbit primary cultivated aortic SMCs and ECs, was shown. In addition, human peripheral monocytes were used and the transmigration of monocytes was observed by scanning electron and laser confocal microscopy. Monocyte differentiation into macrophages was shown by immunohistochemistry and comprehensive gene expression analysis. With the modified form of LDL, the macrophages were observed to accumulate lipids with a foamy appearance and differentiate into the foam cells in the ECM between the ECs and SMCs in the area of our coculture model.