Effects of rabbit antiserum on mouse embryonic stem cells at 8-cell embryo development stage

doi:10.3969/j.issn.0529-1356.2013.06.015

Abstract

Abstract:

Objective Mouse 8-cell embryos were cultured in KSOM media with rabbit antiserum to mouse embryonic stem (ES) cells, and the effect of the antiserum on mouse embryos was observed. Methods The antiserum was diluted to 1/50, 1/100 and 1/200 by KSOM medium, respectively. Results The result of morphology changes showed that the embryonic development was inhibited or even stopped in the 1/50 dilution group, while the compaction was blocked, the embryos showed fluid vacuoles in the blastula stage in the 1/100 dilution group; there was no obvious effect in the 1/200 dilution group. The treatment group with normal rabbit serum did not appear differences from the control group. Our data indicated that the compaction was blocked by antiserum; when the compacted embryos were exposed in the antiserum, decompaction occurred; prolonged decompaction beyond the 32-cell embryo resulted in an increasing proportion of malformed blastocysts. The embryo development was further tested in the 1/100 dilution group by compaction rate, blastocyst rate, inner cell mass (ICM) outgrowth rate, cell counting, embryo transplantation and fluorescent staining. The result showed that the 1/100 dilution antiserum inhibited ICM formation, decreased embryonic cell number, and creased abnormal blastocyst rate. The treated blastocysts were transferred into uterine horns and did not develop to term. Conclusion Our data suggests that the rabbit antiserum to mouse ES cells inhibits embryo compaction, ICM formation and postimplantation development.

Key words: Rabbit antiserum, Embryonic stem cell, Inner cell mass, Compaction, Culture in vitro, Mouse

HU Chun-chao CUI Li-fang WANG Peng-bo WANG Mian-juan DI Ke-qian LI Xiang-yun*. Effects of rabbit antiserum on mouse embryonic stem cells at 8-cell embryo development stage[J]. AAS, 2013, 44(6 ): 812-818.

References

［1］Eakin GS, Hadjantonakis AK, Papaioannou VE, et al. Developmental potential and behavior of tetraploid cells in the mouse embryo［J］. Dev Biol, 2005, 288(1):150-159.
［2］Nagy A, Rossant J, Nagy R, et al. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells ［J］. Proc Natl Acad Sci USA, 1993, 90(18):8424-8428.
［3］Wu XL, Zhang P, Zhao H, et al. Effects of developmental stages of tetraploid embryos on mouse chimeras from embryonic stem cells ［J］. Acta Anatomica Sinica, 2011, 42(5):695-699. (in Chinese)
吴兴龙, 张鹏, 赵虎, 等. 四倍体胚胎发育阶段对胚胎干细胞嵌合体小鼠制备的影响［J］. 解剖学报, 2011, 42(5):695-699. 
［4］Cui XS, Li XY, Shen XH, et al. Transcription profile in mouse four-cell, morula, and blastocyst: genes implicated in compaction and blastocoel formation ［J］. Mol Repord Dev, 2007, 74(2):133-143.
［5］Larue L, Ohsugi M, Hirchenhain J, et al. E-cadherin null mutant embryos fail to form a trophectoderm epithelium［J］. Proc Natl Acad Sci USA, 1994, 91(17):8263-8267.
［6］Butz S, Kemler R. Distinct cadherin-catenin complexesin Ca2+ -dependent cell-cell adhesion［J］. FEBS Lett, 1994, 355(2):195-200.
［7］Piedra J, Martinez D, Castano J, et al. Regulation of β-catenin structure and activity by tyrosine phosphorylation［J］. J Biol Chem, 2001, 276(23):20436-20443.
［8］Ducibella T, Ukena T, Karnovsky M, et al. Changes in cell surface and cortical cytoplasmic organization during early embryogenesis in the preimplantation mouse embryo ［J］. J Cell Biol, 1977,74(1):153-167.
［9］Hinck L, Nathke IS, Papkoff J, et al. Dynamics of cadherin/catenin complex formation: novel protein interactions and pathways of complex assembly［J］. J Cell Biol, 1994,125(6):1327-1340.
［10］Pratt HPM. Lipids and transitions in embryos. In: Johnson MM, eds. Development in Mammals［M］. Vol. 3. Amsterdam: Elsevier, 1978:83-130. 
［11］Johnson MH, Chakraborty J. The effect of prolonged decompaction on the development of the preimplantation mouse embryo ［J］. J Embryol Exp Morph, 1979, 54:241-261.
［12］Cockburn K, Rossant J. Making the blastocyst: lessons from the mouse［J］. J Clin Invest, 2010, 120(4):995-1003.
［13］Artus J, Hadjantonakis AK. Troika of the mouse blastocyst: lineage segregation and stem cells［J］. Curr Stem Cell Res Ther, 2012, 7(1):78-91.
［14］Gasperowicz M, Natale DR. Establishing three blastocyst lineages—then what?［J］. Biol Reprod, 2011, 84(4):621-630.
［15］Hansis C, Grifo J. Oct4 expression in inner cell mass and trophectoderm of human blastocysts［J］. Mol Hum Reprod, 2000, 6(11):999-1004.
［16］Qin PC. Embryology in Mammals［M］. Beijing: Scientific Press, 2001:172-174. (in Chinese)
秦鹏春. 哺乳动物胚胎学［M］. 北京：科学出版社, 2001：172-174. 
［17］Paria BC, Reese J, Das SK, et al. Deciphering the cross-talk of implantation: advance and challenges［J］. Science, 2002, 296(5576):2185-2188.

[1]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[2]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[3]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[4]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[5]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[6]	CHEN Zi-xuan SI Li-na SHU Wei-han ZHANG Xin WEI Chen-yang CHENG Lu-yang YANG Song-he QIAO Yue-bing. Mechanism of melatonin regulation of PI3K/Akt/mTOR signaling in hypothalamus delaying the initiation of puberty in female mice mice [J]. Acta Anatomica Sinica, 2023, 54(6): 644-651.
[7]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.
[8]	LIU Ye CHU Ya-nan XU Ce-lu HE Jia-cheng SU Bing-yin TAI Hao-ran. Roscovitine rescuing neuronal loss and neuroinflammation in brain regions associated with Parkinson’s disease mice [J]. Acta Anatomica Sinica, 2023, 54(6): 635-643.
[9]	WEI Chun-chun WANG Ping LIN Fang-xing MA Xian-hua ZHANG Wei-ping XIE Zhi-fang. An improved fixation method for preparing mouse brown adipose tissue for transmission electron microscopy [J]. Acta Anatomica Sinica, 2023, 54(6): 738-743.
[10]	YAN Kun WU Xiao-lin LIU Ying-juan GE Ke-li1 REN Lei-ming LI Hong-yun . Effect of cerebrotein hydrolysate-Ⅰon intestinal microflora regulation of mice with Parkinson’s disease [J]. Acta Anatomica Sinica, 2023, 54(5): 497-504.
[11]	Ling GU ZHANG Ping SONG KeXin Na Li Hong-yu Chen Zhang Jie CONG Jing ZHAI XiaoYue GU Ling ZHANG Ping SONG Ke-xin LI Na CHEN Hong-yu ZHANG Jie CONG -Jing ZHAI Xiao-yue . Morphometry of the ureteric bud branching in the developing mouse kidney [J]. Acta Anatomica Sinica, 2023, 54(5): 593-598.
[12]	HE Ke-ke LI Yuan-di WEN Ting-hao ZHU Jie GAO Jie HU Rong HAN Feng SU Min . Erythrocyte membrane-associated protein inhibiting Th17 cell differentiation via IL-6/STAT3/ROR-γt signaling pathway in experimental autoimmune encephalomyelitis mice encephalomyelitis mice [J]. Acta Anatomica Sinica, 2023, 54(5): 538-545.
[13]	FAN Wen-juan CHEN Xu-dong CHEN Yong-fang YANG Xu-guang JIN Shao-ju ZHAO Zhi-jun DENG Jin-bo. Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo [J]. Acta Anatomica Sinica, 2023, 54(4): 383-391.
[14]	ZHANG Yi-zhou LI Sha MI Shi-xiong ZUO Hong-chun ZHANG Qian-qian LI Han-lin LEI Zi-han ZHANG Dong-ze CUI Hui-xian. Effect of estradiol regulating sortilin-related receptor A expression on hippocampal spine density and synaptic protein expression via estrogen receptor of mouse [J]. Acta Anatomica Sinica, 2023, 54(3): 261-268.
[15]	LIU Ke-ting CHEN Bin-bin SHUI Yu-lian SHI Qing-ming XIAO Li. Neuroprotective effect of exogenous hydrogen sulfide and the expression of matrix metalloproteinases-9 and Caspase-1 in mice model of Parkinson’s disease [J]. Acta Anatomica Sinica, 2023, 54(3): 289-295.