Figaro Sarah, Pereira Ulysse, Rada Hiram, Semenzato Nicolas, Pouchoulin Dominique, Paullier Patrick, Dufresne Murielle, Legallais Cécile
UMR7338, Laboratory of Biomechanics and Bioengineering, Sorbonne University, University of Technology of Compiègne, CNRS, Compiègne - France.
Gambro Industries, Baxter, Research Center, Meyzieu - France.
Int J Artif Organs. 2017 May 9;40(4):196-203. doi: 10.5301/ijao.5000567. Epub 2017 Mar 21.
Our team previously designed and validated a new bioartificial liver (BAL) called Suppliver based on a Prismaflex™ device, including fluidized bed bioreactors hosting alginate-encapsulated hepatocytes. To ensure correct fluidization within the bioreactor, the beads need to become heavier with the addition of inert glass microspheres.
In this study, we assessed the impact of this additional component on the bead production process, bed fluidization, mass transfer and the mechanical properties of the beads, as well as cell viability and basic metabolic function.
A concentration of 20 mg (1% v/v) of microspheres for 15-20 million cells per milliliter of alginate solution appears to be the best configuration. The filling ratio for the beads in the bioreactors can reach 60%. Four 250-mL bioreactors represent approximately 15% of the hepatocytes in a liver, which is a reasonable target for extracorporeal liver supply.
Increasing bead density clearly maintained the performances of the fluidized bed with plasma of different compositions, without any risk of release out of the bioreactor. A 1% (v/v)-concentration of microspheres in alginate solution did not result in any alteration of the mechanical or biological behavior. This concentration can thus be applied to the production of large-scale encapsulated biomass for further use of the Suppliver setup in human scale preclinical studies.
我们的团队之前设计并验证了一种基于Prismaflex™设备的新型生物人工肝（BAL），即Suppliver，其中包括容纳藻酸盐包封肝细胞的流化床生物反应器。为确保生物反应器内的正确流化，珠子需要通过添加惰性玻璃微球而变得更重。
在本研究中，我们评估了这种额外成分对珠子生产过程、床层流化、传质和珠子机械性能的影响，以及细胞活力和基本代谢功能。
对于每毫升藻酸盐溶液中1500万至2000万个细胞，20毫克（1% v/v）微球的浓度似乎是最佳配置。生物反应器中珠子的填充率可达到60%。四个250毫升的生物反应器约占肝脏中肝细胞的15%，这是体外肝脏供应的合理目标。
增加珠子密度明显维持了不同成分血浆流化床的性能，且不存在从生物反应器中释放出来的风险。藻酸盐溶液中1%（v/v）浓度的微球不会导致机械或生物学行为的任何改变。因此，该浓度可应用于大规模封装生物质的生产，以便在人体规模的临床前研究中进一步使用Suppliver装置。
