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营养供应在造血细胞组织工程生物反应器设计中对细胞生长的作用。

Role of nutrient supply on cell growth in bioreactor design for tissue engineering of hematopoietic cells.

作者信息

Pathi Pragyansri, Ma Teng, Locke Bruce R

机构信息

Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University and Florida A & M University, Tallahassee, Florida 32310-6046, USA.

出版信息

Biotechnol Bioeng. 2005 Mar 30;89(7):743-58. doi: 10.1002/bit.20367.

Abstract

In the present study, a dynamic mathematical model for the growth of granulocyte progenitor cells in the hematopoietic process is developed based on the principles of diffusion and chemical reaction. This model simulates granulocyte progenitor cell growth and oxygen consumption in a three-dimensional (3-D) perfusion bioreactor. Material balances on cells are coupled to the nutrient balances in 3-D matrices to determine the effects of transport limitations on cell growth. The method of volume averaging is used to formulate the material balances for the cells and the nutrients in the porous matrix containing the cells. All model parameters are obtained from the literature. The maximum cell volume fraction reached when oxygen is depleted in the cell layer at 15 days and is nearly 0.63, corresponding to a cell density of 2.25 x 10(8) cells/mL. The substrate inhibition kinetics for cell growth lead to complex effects with respect to the roles of oxygen concentration and supply by convection and diffusion on cell growth. Variation in the height of the liquid layer above the cell matrix where nutrient supply is introduced affected the relative and absolute amounts of oxygen supply by hydrodynamic flow and by diffusion across a gas permeable FEP membrane. Mass transfer restrictions of the FEP membrane are considerable, and the supply of oxygen by convection is essential to achieve higher levels of cell growth. A maximum growth rate occurs at a specific flow rate. For flow rates higher than this optimal, the high oxygen concentration led to growth inhibition and for lower flow rates growth limitations occur due to insufficient oxygen supply. Because of the nonlinear effects of the autocatalytic substrate inhibition growth kinetics coupled to the convective transport, the rate of growth at this optimal flow rate is higher than that in a corresponding well-mixed reactor where oxygen concentration is set at the maximum indicated by the inhibitory kinetics.

摘要

在本研究中,基于扩散和化学反应原理,建立了造血过程中粒细胞祖细胞生长的动态数学模型。该模型模拟了三维(3-D)灌注生物反应器中粒细胞祖细胞的生长和氧气消耗。细胞的物料平衡与三维基质中的营养物质平衡相耦合,以确定传输限制对细胞生长的影响。采用体积平均法来制定含有细胞的多孔基质中细胞和营养物质的物料平衡。所有模型参数均从文献中获取。当细胞层中的氧气在15天时耗尽时,达到的最大细胞体积分数接近0.63,对应细胞密度为2.25×10⁸个细胞/毫升。细胞生长的底物抑制动力学导致了关于氧浓度以及对流和扩散供氧对细胞生长作用的复杂影响。引入营养物质供应的细胞基质上方液层高度的变化,影响了通过流体动力流和跨透气FEP膜扩散的相对和绝对供氧量。FEP膜的传质限制相当大,对流供氧对于实现更高水平的细胞生长至关重要。在特定流速下会出现最大生长速率。对于高于此最佳流速的情况,高氧浓度导致生长抑制,而对于较低流速,由于供氧不足会出现生长限制。由于自催化底物抑制生长动力学与对流传输的非线性效应,在该最佳流速下的生长速率高于相应的充分混合反应器中的生长速率,在该反应器中氧浓度设定为抑制动力学所指示的最大值。

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