Cryopreservation of Taxus chinensis suspension cell cultures.
A simple cryopreservation method for suspension cells of Taxus chinensis was established. In this procedure 7 days old suspension cells were used without any pre-culture treatment. At first, cells were incubated in cryoprotectant solution (0.5M DMSO and 0.5M glycerol) on ice for 30 min and then frozen at a cooling rate of 1 degree C/min to -40 degrees C prior to immersion in liquid nitrogen. The average viability of frozen-thawed cells was between 30 to 40%. The recovery of cryopreserved cells in liquid nitrogen for 1 month was accomplished. After rapid thawing, cells were transferred to solid medium and cultivated for 4-6 weeks. The treatment of trehalose as a cryoprotectant enhanced re-growth of frozen-thawed cells. The stable maintenance of paclitaxel biosynthetic ability in cryopreserved cells was confirmed by comparing with that of regularly sub-cultured suspension cells.
Kim
S I
SI
Samyang Genex Biotech Research Institute,63-2, Hwaam-Dong, Yusung-Gu, Taejon, 305-348, Korea.
Choi
H K
HK
Son
J S
JS
Yun
J H
JH
Jang
M S
MS
Kim
H R
HR
Song
J Y
JY
Kim
J H
JH
Choi
H J
HJ
Hong
S S
SS
D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents; D017694:Rewarming; D020946:Taxus / Q000166:cytology*
D003451:Cryoprotective Agents