| Co-Investigator(Kenkyū-buntansha) |
OOKAWA Keiko Institute of Basic Medical Sciences, Department of Biomedical Engineering, University of Tsukuba, Assistant Professor, 基礎医学系, 講師 (30251052)
OHSHIMA Norio Institute of Basic Medical Sciences, Department of Biomedical Engineering, University of Tsukuba, Professor, 基礎医学系, 教授 (50015971)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
Fetal livers are known to perform hepatic and hematopoietic functions, and contain undifferentiated cells abundantly. In the current study, long-term three-dimensional (3-D) cultures of fetal liver cells (FLCs) as well as bone marrow cells (BMCs) were performed from the tissue engineering viewpoints, and abilities of proliferation and differentiation of these cells were investigated.
As a 3-D scaffold, porous polyvinyl formal (PVF) resin was adopted, and the 3-D cultures as well as conventional monolayer cultures as controls were performed. In the long-term 3-D cultures of murine FLCs for up to 1 month, the FLCs secreted albumin, a representative liver-specific function, more abundantly and stably than those in the monolayer cultures. Moreover, proliferation and albumin secretion of the FLCs were dramatically stimulated by supplementation of oncostatin M, especially in the 3-D cultures.
In the 3-D perfusion cultures of the murine and porcine FLCs, these cells easily lost their viability
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and functions under conditions of ordinary perfusion rate, suggesting that FLCs are quite susceptible to medium flow rates. However, porcine FLCs were rather stable in these perfusion cultures than murine cells, and showed prolonged albumin secretion ability under a low flow rate condition. Therefore, the development of a bioartificial liver using 3-D cultured porcine FLCs is expected to be realistic.
Respecting hematopoietic functions, 3-D cultures of murine BMCs were performed. In the 3-D cultures, high-density cultures of BMCs were achieved, and these cells contained progenitor cells more abundantly than in the monolayer cultures. Moreover, proliferation of the progenitor cells in the 3-D cultures was found to be considerably improved either by coculture of the BMCs with stroma cells or by cultures under a lowered ambient oxygen tension. From these facts, the 3-D cultures of BMCs were considered to be a promising method for effective expansion of the bone marrow immature cells as well. Less
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