Different thawing methods for cryopreserved rabbit limbs: activity of Schwann cells after thawing

doi:10.3969/j.issn.2095-4344.2015.20.016

Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (20): 3200-3204.doi: 10.3969/j.issn.2095-4344.2015.20.016

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Different thawing methods for cryopreserved rabbit limbs: activity of Schwann cells after thawing

Zhu Ze-xing¹, Wang Dan², Zhang Shu-ming¹, Zhao Yan-dong³, Fan Kai-bin⁴, Zhao Yong-jun⁵, Li Hong-jiang⁶, Li Bo⁷, Qiao Lin¹

¹Department of Orthopedics, ²Department of Pharmacology, the Second Artillery General Hospital of Chinese PLA, Beijing 100088, China; ²Fourth People’s Hospital of Linfen City, Linfen 041000, Shanxi Province, China; ⁴Handan Central Hospital, Handan 056001, Hebei Province, China; ⁵Department of Orthopedics, Affiliated Hospital of Logistics University of People’s Armed Police Force, Tianjin 300162, China; ⁶Department of Orthopedics, Tieying Hospital of Fengtai Distirct, Beijing 100078, China; 7Department of Orthopedics, Teda Hospital, Tianjin 300457, China

Online:2015-05-14 Published:2015-05-14
Contact: Qiao Lin, M.D., Associate chief physician, Department of Orthopedics, the Second Artillery General Hospital of Chinese PLA, Beijing 100088, China
About author:Zhu Ze-xing, Master, Attending physician, Department of Orthopedics, the Second Artillery General Hospital of Chinese PLA, Beijing 100088, China

Abstract

Abstract:

BACKGROUND: According to the characteristics of nerve regeneration, the integrity of Schwann cells as regenerative scaffold is very important. Therefore, to save the Schwann cell structure and activity is most important for nerve cryopreservation.
OBJECTIVE: To investigate the optimal thawing procedures for the survival of peripheral nerves in cryopreserved rabbit limbs.
METHODS: Forty hindlimbs from New Zealand White rabbits were randomized into four groups: blank, fast-thawing, slow-thawing and control groups. The control group was subjected to perfusion with protective agent. The severed limbs in fast-thawing and slow-thawing groups were perfused with protective agent followed by gradient cryopreservation and storage in liquid nitrogen for 72 hours. Then, the slow-thawing group was given
gradient thawing procedures, and the fast-thawing group was subjected to fast thawing in 37 ℃ water bath. After thawing, the tibial nerves were harvested for light microscope, electron microscope, immunofluorescence and confocal microscope observation. SPSS17.0 was used for statistical analysis.
RESULTS AND CONCLUSION: No significant difference was found between the blank and control groups. Nerve fiber breakage, axonal damage and mitochondrial swelling were severer in the fast-thawing and slow-thawing groups than the control and blank groups. In addition, the fast-thawing group was superior to the slow-thawing group. These findings indicate that the fast-thawing method has the minimal effects on the survival of nerve fibers in cryopreserved rabbit limbs.

Key words: Replantation, Cryopreservation, Rewarming

CLC Number:

R318

Figures/Tables 5

References

[1] Stachecki JJ,Cohen J,willadsen SM.cryopreservation of unfertilized mouse oocytes:The effect of replacing sodium with choline in the freezing medium．Cryobiology.1998;37(4)：346-354.

[2] Menoret S,Jean M,Tesson L,et al.Optimization of cryopreservation procedures for ratembryos.Transplant Proc. 1999;31(3):1531-1532．

[3] 王增涛,王春霞,朱磊,等.深低温保存81天断指再植1例[J].山东医药,2004,44(3):29.

[4] Jin B,Kusanagi K,Ueda M,et al.Formation of extracellular and intracellular ice during warming of vitrified mouse morulae and its effect on embryo survival.Cryobiology.2008;56(3)：233-240．

[5] Heng BC, Kuleshova LL, Bested SM, et al.The cryopreservation of human embryonic stem cells. Biotechonl Appl Biochem.2005;41(Pt 2):97-104．

[6] Fry LJ,Querol S, Gomez SG,et al. Assessing the toxic effects of DMSO on cord blood to determine exposure time limits and the optimum concentration for cryopreservation.Vox Sang. 2015. doi: 10.1111/vox.12267.

[7] Shimazu T,Mori Y,Takahashi A,et al.Serum-and xeno-free cryopreservation of human umbilical cord tissue as mesenchymal stromal cell source. Cytotherapy. 2015;17(5): 593-600.

[8] Lee JR, Youm HW, Lee HJ,et al. Effect of antifreeze protein on mouse ovarian tissue cryopreservation and transplantation. Yonsei Med J. 2015;56(3):778-784.

[9] Romão R, Marques CC, Baptista MC，et al. Cryopreservation of in vitro-produced sheep embryos: Effects of different protocols of lipid reduction.Theriogenology. 2015. pii: S0093- 691X(15)00105-3.

[10] Tan FC,Lee KH,Gouk SS,et al.Optimization of cryopreservation of stem celIs cultured as neurospheres：comparison between vitrification,slow-cooling and rapid cooling freezing protocols.Cryo Letters.2007;28(6):445-460．

[11] 刘建林,张鹏,黄庆恒.不同复温方法对液氮保存人同种瓣活性影响的实验研究[J].中华实验外科杂志,1999,16(2):112-113.

[12] 瞿玉兴,董天花,张志霖,等.超低温冷冻保存后同种异体神经移植的实验研究[J].中华手外科杂志,2002,18(1):59-62.

[13] Stacy R,Eroglu A,Fowler A,et al.Thermal characterization of Nakagata’s mouse spezm freezing protocol.Cryobiology. 2006; 52(1):99-107．

[14] Milosevic J,Storch A,Schwarz J.Cryopreservation does not affect proliferation and multipotency of murine neural precursor cells.Stem Cells.2005;23(5):681-688．

[15] Dong JP,Malsam J,Bischof JC,et al. Spatial Distrbution of the State of Water in Frozen Mammalian Cell.Biophys J.2010; 99(8):2453-2459.

[16] Vongpralub T,Chinchiyanond W, Hongkuntod P,et al. Cryopreservation of Sambar deer semen in Thailand. Zoo Biol. 2015 Apr 23. doi:10.1002/zoo.21214.

[17] Varela Junior AS,Goularte KL,Alves JP,et al.Methods of cryopreservation of Tambaqui semen, Colossoma macropomum. Anim Reprod Sci. 2015. pii: S0378-4320(15) 00086-X.

Different thawing methods for cryopreserved rabbit limbs: activity of Schwann cells after thawing

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