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IGF-1/IGFBP-1 increases blastocyst formation and total blastocyst cell number in mouse embryo culture and facilitates the establishment of a stem-cell line.

Lin TC, Yen JM, Gong KB, Hsu TT, Chen LR - BMC Cell Biol. (2003)

Bottom Line: We hypothesize that IGF-1 and its dephosphorylated binding protein (IGFBP-1) may be able to improve embryo culture with an associated reduced cell death, and that the resultant increase in the total cell number of the embryo could increase the chances of establishing an embryonic stem-cell line.IGF-1 or dephosphorylated IGFBP-1/IGF-1 supplement does result in an anti-apoptotic effect for early embryo development in culture, with a subsequent increased total cell number resulting from cell culture.The effect is beneficial for the later establishment of a stem-cell line.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gynecology, Obstetrics, and Infertility, Kuo General Hospital, No, 22, Section 2, Ming-Sheng Road, Tainan, 70343, Taiwan. tachin@ksts.seed.net.tw

ABSTRACT

Background: Apoptosis occurs frequently for blastocysts cultured in vitro, where conditions are suboptimal to those found in the natural environment. Insulin-like growth factor-1 (IGF-1) plays an important role in preventing apoptosis in the early development of the embryo, as well as in the progressive regulation of organ development. We hypothesize that IGF-1 and its dephosphorylated binding protein (IGFBP-1) may be able to improve embryo culture with an associated reduced cell death, and that the resultant increase in the total cell number of the embryo could increase the chances of establishing an embryonic stem-cell line.

Results: In vivo fertilized zygotes were cultured in medium containing supplementary IGF-1, or IGFBP-1/IGF-1. The stages of the resultant embryos were evaluated at noon on day five post-hCG injection. The extent of apoptosis and necrosis was evaluated using Annexin V and propidium iodine staining under fluorescent microscopy. The establishment of embryonic stem-cell lines was performed using the hatching blastocysts that were cultured in the presence of IGF-1 or IGFBP-1/IGF-1. The results show that the rate of blastocyst formation in a tissue-culture system in the presence of IGF-1 was 88.7% and IGFBP-1/IGF-1 it was 94.6%, respectively, and that it was significantly greater than the figure for the control group (81.9%). IGFBP-1/IGF-1 also resulted in a higher hatching rate than was the case for the control group (68.8% vs. 48.6% respectively). IGF-1 also increased the number of Annexin V-free and propidium iodine-free blastocysts in culture (86.8% vs. 75.9% respectively). Total cell number of blastocyst in culture was increased by 18.9% for those examples cultured with dephosphorylated IGFBP-1/IGF-1. For subsequent stem-cell culture, the chances of the successful establishment of a stem-cell line was increased for the IGF-1 and IGFBP-1/IGF-1 groups (IGF-1 vs. IGFBP-1/IGF-1 vs. control: 45.8% vs. 59.6% vs. 27.3% respectively).

Conclusion: IGF-1 or dephosphorylated IGFBP-1/IGF-1 supplement does result in an anti-apoptotic effect for early embryo development in culture, with a subsequent increased total cell number resulting from cell culture. The effect is beneficial for the later establishment of a stem-cell line.

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The scheme of conducted experiments
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Figure 1: The scheme of conducted experiments

Mentions: The activity of the IGF system plays a role in embryo development [7,19,22-25]. Kleffens et al. (1999) reported that the expression of IGF system components preceded the occurrence of apoptosis during mouse embryo development and organ-specific development [26]. Further, IGF-1 also prevents the likelihood of induced apoptosis due to the inhibition of caspase-3-like activation and Bcl-2 and Bax expression [13]. Therefore, IGF-1 does play an important role in the prevention of apoptosis in early embryo development and the onward regulation of organ development. We hypothesize that the anti-apoptotic effect of IGF-I and/or its cofactor, IGFBP-1 upon blastomeres, and the reduced likelihood of cell death during blastocyst formation could contribute to an improvement in the results of in vitro embryo culture with reduced cell death rather than the proliferation of cells. Therefore, the possible increased total cell number of embryos in culture with IGF-1 or IGFBP-1/IGF-1 complex could benefit the subsequent embryo culture for the establishment of embryonic stem-cell lines (Figure 1).


IGF-1/IGFBP-1 increases blastocyst formation and total blastocyst cell number in mouse embryo culture and facilitates the establishment of a stem-cell line.

Lin TC, Yen JM, Gong KB, Hsu TT, Chen LR - BMC Cell Biol. (2003)

The scheme of conducted experiments
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC239990&req=5

Figure 1: The scheme of conducted experiments
Mentions: The activity of the IGF system plays a role in embryo development [7,19,22-25]. Kleffens et al. (1999) reported that the expression of IGF system components preceded the occurrence of apoptosis during mouse embryo development and organ-specific development [26]. Further, IGF-1 also prevents the likelihood of induced apoptosis due to the inhibition of caspase-3-like activation and Bcl-2 and Bax expression [13]. Therefore, IGF-1 does play an important role in the prevention of apoptosis in early embryo development and the onward regulation of organ development. We hypothesize that the anti-apoptotic effect of IGF-I and/or its cofactor, IGFBP-1 upon blastomeres, and the reduced likelihood of cell death during blastocyst formation could contribute to an improvement in the results of in vitro embryo culture with reduced cell death rather than the proliferation of cells. Therefore, the possible increased total cell number of embryos in culture with IGF-1 or IGFBP-1/IGF-1 complex could benefit the subsequent embryo culture for the establishment of embryonic stem-cell lines (Figure 1).

Bottom Line: We hypothesize that IGF-1 and its dephosphorylated binding protein (IGFBP-1) may be able to improve embryo culture with an associated reduced cell death, and that the resultant increase in the total cell number of the embryo could increase the chances of establishing an embryonic stem-cell line.IGF-1 or dephosphorylated IGFBP-1/IGF-1 supplement does result in an anti-apoptotic effect for early embryo development in culture, with a subsequent increased total cell number resulting from cell culture.The effect is beneficial for the later establishment of a stem-cell line.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gynecology, Obstetrics, and Infertility, Kuo General Hospital, No, 22, Section 2, Ming-Sheng Road, Tainan, 70343, Taiwan. tachin@ksts.seed.net.tw

ABSTRACT

Background: Apoptosis occurs frequently for blastocysts cultured in vitro, where conditions are suboptimal to those found in the natural environment. Insulin-like growth factor-1 (IGF-1) plays an important role in preventing apoptosis in the early development of the embryo, as well as in the progressive regulation of organ development. We hypothesize that IGF-1 and its dephosphorylated binding protein (IGFBP-1) may be able to improve embryo culture with an associated reduced cell death, and that the resultant increase in the total cell number of the embryo could increase the chances of establishing an embryonic stem-cell line.

Results: In vivo fertilized zygotes were cultured in medium containing supplementary IGF-1, or IGFBP-1/IGF-1. The stages of the resultant embryos were evaluated at noon on day five post-hCG injection. The extent of apoptosis and necrosis was evaluated using Annexin V and propidium iodine staining under fluorescent microscopy. The establishment of embryonic stem-cell lines was performed using the hatching blastocysts that were cultured in the presence of IGF-1 or IGFBP-1/IGF-1. The results show that the rate of blastocyst formation in a tissue-culture system in the presence of IGF-1 was 88.7% and IGFBP-1/IGF-1 it was 94.6%, respectively, and that it was significantly greater than the figure for the control group (81.9%). IGFBP-1/IGF-1 also resulted in a higher hatching rate than was the case for the control group (68.8% vs. 48.6% respectively). IGF-1 also increased the number of Annexin V-free and propidium iodine-free blastocysts in culture (86.8% vs. 75.9% respectively). Total cell number of blastocyst in culture was increased by 18.9% for those examples cultured with dephosphorylated IGFBP-1/IGF-1. For subsequent stem-cell culture, the chances of the successful establishment of a stem-cell line was increased for the IGF-1 and IGFBP-1/IGF-1 groups (IGF-1 vs. IGFBP-1/IGF-1 vs. control: 45.8% vs. 59.6% vs. 27.3% respectively).

Conclusion: IGF-1 or dephosphorylated IGFBP-1/IGF-1 supplement does result in an anti-apoptotic effect for early embryo development in culture, with a subsequent increased total cell number resulting from cell culture. The effect is beneficial for the later establishment of a stem-cell line.

Show MeSH
Related in: MedlinePlus