Cryopreservation of Panax ginseng cells |
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Authors: | A. Joshi W.-L. Teng |
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Affiliation: | (1) Hong Kong Institute of Biotechnology Ltd., 2 Biotechnology Ave., 12 Miles Tai Po Road, Shatin, N.T., Hong Kong Fax: +852-26036820 e-mail: wl-teng@hkib.org.hk, HK |
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Abstract: | The impacts of cryoprotectants (CP) and cell status during the growth cycle on Panax ginseng cell viability during cryopreservation were investigated. The ginseng cells used had a 5–7 times proliferation rate (compared with inoculum) in 2–3 weeks and were subcultured at 2- and 4-week intervals in liquid and on solid media, respectively. After testing various CP solutions of glycerol, dimethylsulphoxide, ethylene glycol and sucrose, a combination of 10% (v/v) glycerol and 4% (w/v) sucrose was selected for its least cytotoxicity and highest cell viability after thawing. With this CP solution, cells throughout the growth cycle exhibited a ‘U’-shaped fluctuation of post-thaw cell viability. The highest viability (86.5%) occurred during the lag phase from cells already maintained in suspension culture and then in the late exponential phase (61.7%); the lowest level of 15.4% was in the mid-exponential phase. Callus freshly transferred to liquid medium showed a less obvious fluctuation pattern. The recovered cells were brown-to-reddish at first and gradually returned to a light yellow colour after several subcultures. Received: 1 October 1998 / Revision received: 6 January 2000 / Accepted: 11 January 2000 |
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Keywords: | Panax ginseng Suspension Cryopreservation Rapid freezing |
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