Comparison of methods to isolate and cryopreserve goose primordial germ cells

doi:10.3969/j.issn.1004-1524.2017.03.08

›› 2017, Vol. 29 ›› Issue (3): 401-407.DOI: 10.3969/j.issn.1004-1524.2017.03.08

• Animal Science • Previous Articles Next Articles

Comparison of methods to isolate and cryopreserve goose primordial germ cells

YU Jianning, YAN Leyan, CHEN Zhe, SHAO Xibing, DAI Zichun, YING Shijia, SHI Zhendan^*

Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Laboratory of Animal Improvement and Reproduction, Nanjing 210014, China

Received:2016-11-23 Online:2017-03-20 Published:2017-03-31

Abstract

Abstract: In order to establish the isolation and cryopreservation method for goose primordial germ cells (PGCs), four kinds of PGCs separation methods,which were blood Ficoll density gradient centrifugation, blood ACK lysis method, gonads trypsin-EDTA digestion method and gonads digestion combined with short-term culture method, were compared in this experiment. And three kinds of cryoprotectors that were STEM-CELLBANKER, CELLBANKER 2 and 10% DMSO plus 90% serum were compared to evaluate the cryopreservation efficiency of goose PGCs. Morphological observation, PAS and PGCs specific immunofluorescence staining were used to verify PGCs, and trypan blue staining was used to detect the vitality of PGCs. The results showed that goose PGCs were successfully isolated from the blood and gonad of goose embryo. Gonads digestion combined with short-term culture method got the highest goose PGCs quantity, purity and vitality, reached 115 cells per embryo, 80.8% and 88.3% respectively, compared with the traditional blood separation method. Commercial stem-cell cryoprotector STEM-CELLBANKER was more suitable for cryopreservation of goose PGCs than the normal cell cryoprotectants, and the cell survival rate was about 90% after thawing. Gonads digestion combined with short-term culture method could get sufficient quantity and purity of goose PGCs. STEM-CELLBANKER could be used for the long-term cryopreservation of goose PGCs.

Key words: goose, primordial germ cells, gonads digestion combined with short-term culture, cryopreservation

CLC Number:

S835

YU Jianning, YAN Leyan, CHEN Zhe, SHAO Xibing, DAI Zichun, YING Shijia, SHI Zhendan. Comparison of methods to isolate and cryopreserve goose primordial germ cells[J]. , 2017, 29(3): 401-407.

References

[1] 汤青萍,章双杰,郭军,等. 太湖鹅群体遗传多样性研究[J]. 家畜生态学报,2010,31(1):30-33.
TANG Q P, ZHANG S J, GUO J, et al. Analysis of genetic diversity in Taihu geese[J]. Journal of Domestic Animal Ecology , 2010, 31(1):30-33. (in Chinese with English abstract)
[2] 吴常信. 畜禽遗传资源保存的理论与技术[J]. 家畜生态,2001,22 (1):1-4.
WU C X. Theory and technology of preservation in domestic animal genetic resources[J]. Ecology of Domestic Animal , 2001, 22 (1):1-4. (in Chinese with English abstract)
[3] NAKAMURA Y, TASAI M, TAKEDA K, et al. Production of functional gametes from cryopreserved primordial germ cells of the Japanese quail[J]. Journal of Reproduction and Development , 2013, 59(6):580-587.
[4] NAKAMURA Y, USUI F, MIYAHARA D, et al. Efficient system for preservation and regeneration of genetic resources in chicken: concurrent storage of primordial germ cells and live animals from early embryos of a rare indigenous fowl (Gifujidori)[J]. Reproduction Fertility and Development , 2010, 22(8):1237-1246.
[5] NAKAMURA Y, KAGAMI H, TAGAMI T. Development, differentiation and manipulation of chicken germ cells[J]. Development , Growth and Differentiation , 2013, 55(1):20-40.
[6] 韩毅冰,于康震,周琦,等. 鸡胚血液中原始生殖细胞的分离及其培养的研究[J]. 生物技术,1996,6(2):11-13.
HAN Y B, YU K Z, ZHOU Q, et al. Studies on the isolation and culture of primordial germ cells from blood of chicken embryos[J]. Biotechnology , 1996, 6(2):11-13. (in Chinese)
[7] SONG Y, DURAISAMY S, ALI J, et al. Characteristics of long-term cultures of avian primordial germ cells and gonocytes[J]. Biology of Reproduction , 2014, 90(1):15.
[8] 李碧春,秦洁,肖小琚,等. 鸡胚不同发育时期原始生殖细胞的分离方法[J]. 动物学报,2003,49(6):835-842.
LI B C, QIN J, XIAO X X, et al. A method for isolation of primordial germ cells in different stages of the chicken embryo[J]. Acta Zoologica Sinica , 2003, 49(6):835-842. (in Chinese with English abstract)
[9] YAMAMOTO Y, USUI F, NAKAMURA Y, et al. A novel method to isolate primordial germ cells and its use for the generation of germline chimeras in chicken[J]. Biology of Reproduction , 2007, 77(1): 115-119.
[10] 李赞东,刘春海,黄劲松,等. 鸡鸭异种间嵌合体的制备[J]. 动物学报,2002,48(4):543-548.
LI Z D, LIU C H, HUANG J S, et al. Preparation of Interspecies chicken ( Gallus domesticus )/duck ( Anas domesticus ) chimeras[J]. Acta Zoologica Sinica , 2002, 48(4):543-548. (in Chinese with English abstract)
[11] ZHANG W, RUI L, ZHANG J, et al. Production of chimeras between the Chinese soft-shelled turtle and Peking duck through transfer of early blastoderm cells[J]. Journal of Experimental Biology , 2013, 216(Pt 10):1786-1792.
[12] YAN L, GAO J S, RUI L, et al. A review of strategies for producing chimeric birds[J]. Avian Biology Research , 2014, 7(1):3-9.
[13] 孙鹏翔. 鸡胚原始生殖细胞体外转染及其介导的转基因研究[D]. 扬州:扬州大学,2007:19-32.
SUN P X. Study on the in vitro transfection and transgenic with embryonic primordial germ cells [D]. Yangzhou: Yangzhou University, 2007:19-32. (in Chinese with English abstract)
[14] 唐新燕,张才乔. 鸡胚原始生殖细胞体外的分化[J]. 农业生物技术学报,2007,15(1):20-23.
TANG X Y, ZHANG C Q. Differentiation of chicken primordial germ cells in vitro[J]. Journal of Agricultural Biotechnology , 2007, 15 (1):20-23. (in Chinese with English abstract)
[15] 葛楚天. 鸡胚原始生殖细胞发育调控及其机理的研究[D]. 杭州:浙江大学,2010:38.
GE C T. Regulation of the chicken primordial germ cell development [D]. Hangzhou: Zhejiang University, 2010:38. (in Chinese with English abstract)

Comparison of methods to isolate and cryopreserve goose primordial germ cells

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