PEPTIDES EXTRACTED FROM VERO CELL-CULTURES OVERCOME THE BLASTOCYST BLOCK OF MOUSE EMBRYOS IN A SERUM-FREE MEDIUM

被引：18

作者：

CHEN, HF ^{[1
]}

HO, HN ^{[1
]}

CHEN, SU ^{[1
]}

CHAO, KH ^{[1
]}

LIN, HR ^{[1
]}

HUANG, SC ^{[1
]}

LEE, TY ^{[1
]}

YANG, YS ^{[1
]}

机构：

[1] NATL TAIWAN UNIV,COLL MED,DEPT OBSTET & GYNECOL,TAIPEI,TAIWAN

来源：

JOURNAL OF ASSISTED REPRODUCTION AND GENETICS | 1994年 / 11卷 / 03期

关键词：

PEPTIDES; COCULTURE; VERO CELL; BLASTOCYST BLOCK; MOUSE EMBRYO;

D O I：

10.1007/BF02332095

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Purpose: The aim of this work was to evaluate the effect of a Vero cell coculture system on the development of mouse embryos. Methods: Mouse embryos were randomly divided and cultured in human tubal fluid (HTF) medium with/without Vero cell monolayers, conditioned medium (CM) obtained from Vero cell cultures, and HTF medium supplemented with peptides extracted from CM. The concentrated CM was examined by SDS/PAGE. Results: The development of mouse embryos was blocked at the blastocyst stage in pur HTF medium (1.4% hatching at day 5). This ''blastocyst block'' was overcome by coculture with Vero cell monolayers (48.1% hatching at day 5; 1.4 vs 48.1%; P < 0.001). CM and the addition of 5% fetal bovine serum (24.1 and 34.9% hatching, respectively, at day 5) were also able to enhance the process of hatching. In the other experiment, the addition of peptides extracted from Vero cell cultures also overcame the blastocyst block (12.5%) compared with pure HTF medium (2.1%) (P < 0.05). Electrophoretic separation revealed several classes of polypeptides consistently secreted into CM obtained from Vero cell cultures. Most peptides occurred in the M(r) range between 6.5 kd and 35.9 kd. Conclusion: A developmental block (blastocyst block) of mouse embryos in a serum- and protein-free medium (HTF) was discovered in this study. This block was effectively overcome by HTF plus serum and coculture with Vero cell monolayers and also by the peptides extracted from Vero cell-conditioned medium. We speculate that certain factors secreted or converted by Vero cells may be critical in hatching of mouse embryos. Further study of these factors may be helpful in delineating its mechanism.

引用

页码：165 / 171

页数：7

共 20 条

[1]

Edwards R.G., Steptoe P.C., Purdy J.M., Establishing full term human pregnancies using cleaving embryos grown in vitro, Br J Obstet Gynecol, 87, pp. 737-740, (1980)

[2]

In vitro fertilization-embryo transfer (IVF-ET) in the United States: 1990 results from the IVF-ET Registry, Fertil Steril, 57, pp. 15-24, (1992)

[3]

Bongso A., Ng S.C., Mok H., Lim M.N., Teo H.L., Wong P.C., Ratnam S.S., Effect of sperm motility on human in vitro fertilization, Arch Androl, 22, pp. 185-190, (1989)

[4]

Bolton V.N., Hawes S.M., Taylor C.T., Parsons J.H., Development of spare human preimplantation embryos in vitro: An analysis of the correlations among gross morphology, cleavage rates, and development to the blastocyst, J Vitro Fert Embryo Transfer, 6, pp. 30-35, (1989)

[5]

Bongso A., Ng S.C., Fong C.Y., Ratnam S., Cocultures: A new lead in embryo quality improvement for assisted reproduction, Fertil Steril, 56, pp. 179-191, (1991)

[6]

Ouhibi N., Hamidi J., Guillaud J., Menezo Y., Co-culture of 1-cell mouse embryos on different cell supports, Hum Reprod, 5, pp. 737-743, (1990)

[7]

Czlonkowska M., Eysymont U., Guszkiewicz A., Kossakowski M., Dziak J., Birth of lambs after in vitro maturation, fertilization, and coculture with oviductal cells, Mol Reprod Dev, 30, pp. 34-38, (1991)

[8]

Goldberg J.M., Khalifa E.A.D.M., Friedman C.I., Kim M.H., Improvement of in vitro fertilization and early embryo development in mice by coculture with human fallopian tube epithelium, Am J Obstet Gynecol, 165, pp. 1802-1805, (1991)

[9]

Eyestone W.H., First N.L., Co-culture of early cattle embryos to the blastocyst stage with oviductal tissue or in conditioned medium, J Reprod Fertil, 85, pp. 715-720, (1980)

[10]

Ellington J.E., Carney E.W., Farrell P.B., Simkin M.E., Foote R.H., Bovine 1-2-cell embryo development using a simple medium in three oviduct epithelial cell coculture systems, Biol Reprod, 43, pp. 97-104, (1990)

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