F-0725 高細胞密度人工組織の凍結保存後生存率に及ぼす冷却速度の影響(J22-4 生体関連の熱・流体工学(4))(J22 生体関連の熱・流体工学)

概要

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The effect of cooling rate on the post-thaw viability of cells in cryopreserved artificial tissue was studied. Human fibroblasts were three-dimensionally cultured for 2 days in collagen sponge (φ20x1mm) as an extracellular matrix to imitate biological tissue (artificial tissue). Different cell densities for the artificial tissue were used, from 10^5 to 10^7 cells/cm^3. Four artificial tissues were first stacked in a test chamber, then frozen at a cooling rate from 0.3 to 50℃/min in a solution of Dulbecco's Modified Eagle Medium, 20% Fetal Bovine Serum, and 10% dimethylsulfoxide, then kept frozen at -196℃ for 2 hours, and finally thawed. Collagen matrix of artificial tissue was resolved by using collagenase. Post-thaw viability of fibroblasts was evaluated by using a trypan blue exclusion assay. Results show that with increasing cell density, the post-thaw viability decreased. Moreover, when the cell density was high, dehydration was obstructed, and thus the optimal cooling rate became slower.
一般社団法人日本機械学会の論文
2001-08-22