| Co-Investigator(Kenkyū-buntansha) |
KIMURA Minoru TokaiUniversity, School of Medicine, Professor, 医学部, 教授 (10146706)
NAKAZAWA Hiroe TokaiUniversity, School of Medicine, Professor, 医学部, 教授 (20110885)
KAKUTA Takatoshi TokaiUniversity, School of Medicine, Assistant Professor, 医学部, 講師 (50276854)
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| Research Abstract |
Current hemodialysis is remarkably insufficient as an artificial kidney because it is an intermittent treatment using7.2% of 168 hours of a week for dialysis and does not have tubular function with metabolic and endcrinologic roles. Treatment cost of hemodialysis is increasing year by year not only for enlarged total number of dialysis patients, but also for large number of patients with severe complications. We intended to develop a bioartificial kidney in which 6 litters(L) of 10L/day of continuous ultrafiltrate were regenerated by artificial tubules, substituting 4L/day with meal and drinks, since we had confirmed urea, creatinine and β_2 microglobulin were maintained at very low levels by continuous hemofiltration with 10L/day of filtrate. Selection of suitable kind of tubular epithelial cells, artificial membranes, determination of composition of culture medium, method for medium distribution, seeding method of the cells and evaluation method of tubular functions were studied as t
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he fundamental investigation for development of bioartificial tubules. As the result of the study, LLC-PK_1,cells, porcine proximal tubular epithelial cells, were seeded inside the polysulfone hollow fibers coated with type I collagen on the inner surface. The seeding was done four times-at every 90 of module rotation with 10^8 cells/ml. Confluent monolayers of LLC-PK_1, cells were successfully formed with this condition regardless of width of membrane area of hollow fibers. We evaluated transport abilities of H_2O, Na^+ and glucose, and leak rates of urea and creatinine of the bioartificial tubules by perfusion of medium with 50mg/dl of urea nitrogen and 5.0mg/dl of creatinine inside the hollow fibers. H_2O, Na^+ and glucose transport by artificial tubules with 1m^2 of membrane area for 24 hours were 67, 93, 70% of the targeted values of 6L of 10L/day of filtrate, respectively. Membrane area, 1.3m^2 of bioartificial tubules is needed for the transport of 6L of daily filtrate. Compact, wearable artificial tubules would be developed using aquaporin 1 transfected tubular epithelial cells, since water transport ability of rat aquaporinl DNA transfected LLC-PK_1, cells was about two-fold of that of wild type LLC-PK_1, cells. Transfection of genes of other functional proteins into tubular epithelial cells must be investigated for development of bioartificial tubules. Less
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