Evaluation of Viability of Cryopreserved Rat Cardiac Myocytes and Effects of Dimethyl Sulfoxide Concentration on Cryopreservation

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  • ラット心筋細胞の凍結保存におけるジメチルスルホキシドの有効性評価
  • ラット シンキン サイボウ ノ トウケツ ホゾン ニ オケル ジメチルスルホキシド ノ ユウコウセイ ヒョウカ

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Abstract

Cryopreserved cardiac myocytes are expected to be useful for the development of new drug against heart disease. However, the appropriate cryopreservation process for cardiac myocytes has not been established yet. Therefore, in this study, we evaluate the effectiveness of dimethyl sulfoxide (DMSO) on the cryopreservation of cardiac myocytes by three measurements: 1) the total viability (including cardiac myocytes) just after thawing with trypan blue staining, 2) the observation of morphology of cardiac myocytes for 7 days culture by real-time phase contrast microscopy and immunofluorescent staining, and 3) the variation of their beating rates for 7 days after thawing. The total viabilities of cryopreserved cells with 3v/v% to 20v/v% DMSO concentration were higher than those without DMSO. Thawed cells contracted regularly and spontaneously during culture, as did the non-cryopreserved cells. The immunofluorescence observation indicated that the thawed cardiac myocytes expressed α-actinin. Therefore, cardiac myocytes could be survived by the cryopreservation with DMSO concentrations between 3v/v% and 20v/v%, which had the normal contraction rates during culture for 7 days.

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