• Journal of Embryo Transfer
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• v.31 no.3
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• pp.169-177
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• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• Geotechnical Engineering
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• v.12 no.4
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• pp.157-178
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• 1996

Current design methods for reinforced earth structures take no account of the magnitude of the strains induced in the tensile members as these are invariably manufactured from high modulus materials, such as steel, where straits are unlikely to be significant. With fabrics, however, large strains may frequently be induced and it is important to determine these to enable the stability of the structure to be assessed. In the present paper internal design method of analysis relating to the use of fabric reinforcements in reinforced earth structures for both stress and strain considerations is presented. For the internal stability analysis against rupture and pullout of the fabric reinforcements, a strain compatibility analysis procedure that considers the effects of reinforcement stiffness, relative movement between the soil and reinforcements, and compaction-induced stresses as studied by Ehrlich 8l Mitchell is used. I Bowever, the soil-reinforcement interaction is modeled by relating nonlinear elastic soil behavior to nonlinear response of the reinforcement. The soil constitutive model used is a modified vertsion of the hyperbolic soil model and compaction stress model proposed by Duncan et at., and iterative step-loading approach is used to take nonlinear soil behavior into consideration. The effects of seepage pressures are also dealt with in the proposed method of analy For purposes of assessing the strain behavior oi the fabric reinforcements, nonlinear model of hyperbolic form describing the load-extension relation of fabrics is employed. A procedure for specifying the strength characteristics of paraweb polyester fibre multicord, needle punched non-woven geotHxtile and knitted polyester geogrid is also described which may provide a more convenient procedure for incorporating the fablic properties into the prediction of fabric deformations. An attempt to define improvement in bond-linkage at the interconnecting nodes of the fabric reinforced earth stracture due to the confining stress is further made. The proposed method of analysis has been applied to estimate the maximum tensions, deformations and strains of the fabric reinforcements. The results are then compared with those of finite element analysis and experimental tests, and show in general good agreements indicating the effectiveness of the proposed method of analysis. Analytical parametric studies are also carried out to investigate the effects of relative soil-fabric reinforcement stiffness, locked-in stresses, compaction load and seepage pressures on the magnitude and variation of the fabric deformations.