Extracorporeal Bioartificial Liver Using the Radial‐flow Bioreactor in Treatment of Fatal Experimental Hepatic Encephalopathy

Kanai, Hideki; Marushima, Hideki; Kimura, Naofumi; Iwaki, Takamasa; Saito, Masaya; Maehashi, Haruka; Shimizu, Keiko; Muto, Makiko; Masaki, Takahiro; Ohkawa, Kiyoshi; Yokoyama, Keitaro; Nakayama, Masaaki; Harada, Tohru; Hano, Hiroshi; Hataba, Yoshiaki; Fukuda, Takahiro; Nakamura, Masahiko; Totsuka, Naoto; Ishikawa, Shutaro; Unemura, Yasuki; Ishii, Yuji; Yanaga, Katsuhiko; Matsuura, Tomokazu

doi:10.1111/j.1525-1594.2007.00354.x

Cited by 28 publications

(19 citation statements)

References 6 publications

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“…Culture conditions were employed as reported previously (20,36,37). Briefly, A431 cells (1x10 7 cells) that had been injected in the reservoir of the RFB system were trapped by adherence to the matrix and then continuously cultured with 150 ml of RPMI-1640 medium supplemented 10% FBS in a CO 2 incubator (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…In contrast to CA9 protein expression under hypoxia in the living body, in vitro regulation of the protein expression is reported to be positively related to dense culture (78,79). In order to determine whether or not CA9 molecular induction in 3D cultured cells was dependent on the increasing cell density, the effects of cell density on CA9 protein expression levels were analyzed using a 2D, monolayer culture dish, the RFB and other culture devices to provide spheroid culture conditions, since 3D culture in the RFB has been known to culture cells at a high density while mimicking the in vivo architecture (37,80). Results showed that the increased protein levels of CA9 at various degrees under 3D culture conditions in all devices yielded cell densities at lower than 50% confluence and showed non-significant cell density-dependent induction of CA9 mRNA expression during the period of the experiment in the 2D cultured cells (Fig.…”

Section: Increased Expression Of Ca9 In Cultured Cells Under 3dmentioning

confidence: 99%

“…Examination of the molecular and biological mechanisms of the cancer cell can be carried out in the environment using a 3D culture system that mimics the living organism, mainly through a spheroid culture system. Recent progress has been demonstrated with many types of bioreactor for 3D and highdensity cell culture systems (20,36,37). In our laboratory, a radial flow type bioreactor (RFB) was developed and several characteristics of RFB-cultured cells were determined.…”

Section: Introductionmentioning

confidence: 99%

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Three-dimensional culture promotes reconstitution of the tumor-specific hypoxic microenvironment under TGFβ stimulation

Marushima

Shibata²,

Asakura³

et al. 2011

Int J Oncol

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Abstract.In vitro tumor growth in a three-dimensional (3D) architecture has been demonstrated to play an important role in biology not only for developmental organogenesis and carcinogenesis, but also for analyses on reconstitution and maintenance in a variety of biological environments surrounding the cells. In addition to providing architectural similarity to living organisms, 3D culture with a radial flow bioreactor (RFB) can also closely mimic the living hypoxic microenvironment under which specific organogenesis or carcinogenesis occurs. The findings of the present study under the RFB culture conditions show that cancer cells underwent a shift from aerobic to hypoxic energy metabolism, in addition to protein expression to maintain the 3D structure. In RFB-cultured cells, protein stability of hypoxia-inducible factor 1 (HIF1) α, a subunit of HIF1, was increased without upregulation of its mRNA. Under these conditions, PHD2, HIF-prolyl-4-hydroxylase 2 and a HIF1 downstream enzyme, were stabilized without affecting the mRNA levels via downregulation of FK506-binding protein 8. PHD2 accumulation, which occurred concomitant with HIF1 stabilization, may have compensated for the lack of oxygen under hypoxic conditions to regulate the HIF levels. 3D-culture-induced overexpression of carbonic anhydrase (another representative HIF downstream enzyme) was found to occur independently of cell density in RFBcultured cells, suggesting that the RFB provided an adequately hypoxic microenvironment for the cultured cells. From these results, it was hypothesized that the key factors are regulatory molecules, which stabilize and degrade HIF molecules, thereby activating the HIF1 pathway under a hypoxic milieu.

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Section: Methodsmentioning

confidence: 99%

Section: Increased Expression Of Ca9 In Cultured Cells Under 3dmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Three-dimensional culture promotes reconstitution of the tumor-specific hypoxic microenvironment under TGFβ stimulation

Marushima

Shibata²,

Asakura³

et al. 2011

Int J Oncol

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“…При наличии пористого носителя (подложки), гепатоциты могут функционировать внутри него, при этом они защищены от раневого стресса, что позволяет использовать более высокие скорости перемешивания и аэрации в процессе культивирования. В этом качестве были исследованы различные материалы, в том числе полиуретановая пена [95], поливиниловая смола [96], альгинатные частицы [97], пористый гидроксиапатит [98] и полиэфирные скаффолды [99].…”

Section: системы с "уплотнённым слоем" (Packed Bed System)unclassified

Problems and prospects of creation of extracorporal systems for support of functional livers status

2015

BIOMED KHIM

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Рассмотрены особенности эфферентной терапии с использованием экстракорпоральных систем, аппаратов "искусственной печени" и "биоискусственной печени" при лечении печёночной недостаточности. Анализ литературных данных показывает необходимость дальнейшего развития этих биомедицинских исследований и поиска оптимальных решений для подбора источника гепатоцитов, разработки биореакторов и биоматериалов, составляющих основу аппаратов типа "Биоискусственная печень". Учет определенных преимуществ и недостатков различных методов экстракорпоральной поддержки функционального состояния печени позволяет оценить предшествующий опыт при лечении заболеваний печени и подойти к разработке новых, более эффективных лечебных технологий.Ключевые слова: биоискусственная печень, гепатоциты, биоматериалы, печёночная недостаточность DOI: 10.18097/PBMC20156105545 предшествующий опыт при лечении заболеваний печени, но и подойти к разработке новых, более эффективных лечебных технологий.

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“…In our laboratory, several cell lines derived from hepatocellular carcinoma culture in a radial flow type bioreactor (RFB), which is a type of 3D culture system with a high density cell population, showed not only improved cell functions, but also a differentiated morphology as compared with those grown in 2D culture (5,(9)(10)(11). A representative result demonstrating the functional effectiveness of this system was that use of the RFB system to culture functional liver cells cured a mini-pig with amanitin-, galactosamine-induced acute liver failure as it served as an artificial liver system (10).…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway

Ohkawa¹

2009

Int J Oncol

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Abstract.To confirm the usefulness of the radial flow type bioreactor (RFB) for a three-dimensional (3D) culture system, which provides a tissue architecture and molecular function mimicking the in vivo environment, molecular expression in the A431 human squamous carcinoma cell line during culture were analyzed under the physically different environments of 3D culture in the RFB, 2D culture in a monolayer as well as in nude mice. Time-dependent accumulation of autocrine transforming growth factor (TGF) ß1 was found in spent culture media obtained only from 3D cultured A431 cancer cells, which grew well with a stratified-sheet morphology. Cells in the RFB overexpressed matrix metalloproteinase 7 (MMP7) and showed an increased release of soluble 80-kDa fragments of E-cadherin into the media time-dependently, resulting in the reduction of E-cadherin protein at the cell surface without down-regulation of the mRNA. ß-Catenin and its nuclear partner, LEF1, were up-regulated and Wnt protein secretion was also accelerated. Additional up-regulation of the transcriptional factors, HMGA2 and down-stream Slug, was noted. TGFß1-dependent, MMP7-mediated up-regulation of ß-catenin/LEF1 signaling and TGFß1-activated HMGA2 pathways consequently converged with Slug overexpression, due to disassembly and further repression of E-cadherin expression, which was reproducible in the epithelial mesenchymal transition process without any manipulation. Other transcriptional factors, Notch/HEY1 and NF-κB, were also up-regulated in 3D-cultured cells. These signals recruited molecules related to extracellular matrix-cell remodeling and angiogenesis. Expression of several representative molecules in the 3D cultured cells was parallel with that in xenotransplanted A431 tumor tissues in nude mice. 3D culture of tumor cells in the RFB is a useful tool for cancer experimental biology and evaluation of cancer therapeutic-like systems in nude mice.

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Extracorporeal Bioartificial Liver Using the Radial‐flow Bioreactor in Treatment of Fatal Experimental Hepatic Encephalopathy

Cited by 28 publications

References 6 publications

Three-dimensional culture promotes reconstitution of the tumor-specific hypoxic microenvironment under TGFβ stimulation

Three-dimensional culture promotes reconstitution of the tumor-specific hypoxic microenvironment under TGFβ stimulation

Problems and prospects of creation of extracorporal systems for support of functional livers status

Three-dimensional culture using a radial flow bioreactor induces matrix metalloprotease 7-mediated EMT-like process in tumor cells via TGFβ1/Smad pathway

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