The effect of preimplantation culture conditions on murine embryo implantation and fetal development

Arny, M.; Nachtigall, Lila E.; Quagliarello, John

doi:10.1016/s0015-0282(16)59545-4

Cited by 44 publications

(15 citation statements)

References 20 publications

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“…More embryos cultured in KSOM supplemented with FCS died before they reached the blastocyst stage. In agreement with our results, serum has been found to reduce the postimplantation viability of embryos (20,26,27). Blastocysts that were cultured in the presence of FCS showed a normal appearance; however, a higher proportion of them did not survive after transfer into recipient females compared with blastocysts obtained in the control group.…”

Section: Discussionsupporting

confidence: 91%

Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior

Fernández‐González

Moreira

Bilbao

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

354

271

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The long-term developmental and behavioral consequences of mammalian embryo culture are unknown. By altering the culture medium with the addition of FCS, we wanted to determine whether mouse embryos cultured under suboptimal conditions develop aberrant mRNA expression of imprinting genes at the blastocyst stage and whether fetal development, growth, and behavior of adult mice are affected. One-cell embryos obtained from superovulated female B6CBAF1 mice were cultured for 4 days in K ؉ -modified simplex optimized medium in the presence of either 10% FCS or 1 g͞liter BSA. After embryo transfer, born animals were submitted to several developmental and behavior tests. The mRNA expression of some imprinting genes was significantly affected in blastocysts cultured in the presence of FCS. Two of the eight measures of preweaning development and some specific measures of neuromotor development, such as the walking activity, were delayed in the group originated with FCS. After 34 weeks, the weight of female mice cultured in vitro in the presence of FCS was significantly higher than controls. In addition, the locomotion activity of mice was altered at 5 and 15 months. Anatomopathological and histological analysis of animals at 20 months of age showed some large organs and an increase in pathologies. We have found that mice derived from embryos cultured with FCS exhibited specific behavioral alterations in anxiety and displayed deficiencies in implicit memories. Our data indicate that long-term programming of postnatal development, growth, and physiology can be affected irreversibly during the preimplantation period of embryo development by suboptimal in vitro culture.embryo culture ͉ organ weight ͉ postnatal defects

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Section: Discussionsupporting

confidence: 91%

Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior

Fernández‐González

Moreira

Bilbao

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

354

271

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“…We observed a reduced implantation rate in those groups in which embryos were cultured in vitro to the blastocyst stage, suggesting a negative relationship between ICSI and in vitro culture. This negative effect has been previously reported (Arny et al, 1987;Van der Auwera et al, 1999;Khosla et al, 2001a;Khosla et al, 2001b;Ferná ndez-Gonzalez et al, 2004;Pérez-Crespo et al, 2005), and it questions the convenience of culturing embryos to the blastocyst stage to allow the selection of chromosomally competent embryos that has been proposed by other authors (Sakkas, 1999).…”

Section: Discussionmentioning

confidence: 81%

Factors From Damaged Sperm Affect Its DNA Integrity and Its Ability to Promote Embryo Implantation in Mice

Pérez-Crespo

Moreira

Pintado

et al. 2008

Journal of Andrology

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Endogenous nucleases in mouse sperm can be activated by freeze-thawing the spermatozoa in media without cryoprotection and cleaving spermatozoa DNA. The role of sperm chromatin integrity during intracytoplasmic sperm injection (ICSI) is of critical importance. We analyzed in the B6D2 mouse the proportion of DNA-fragmented spermatozoa (DFS) produced by incubation in conditioned medium (CM) generated by freeze-thawing sperm in the absence of cryoprotection. We then examined the subsequent development, implantation, and offspring obtained after ICSI with incubated spermatozoa. When fresh sperm cells were incubated for 90 minutes in this CM, a significant increase in the amount of DFS was detected by the terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay (27% vs 4.5% in fresh sperm). After ICSI of fresh and incubated spermatozoa, embryos were cultured in vitro to either the 2-cell or blastocyst stage before they were transferred into pseudopregnant CD1 females. On day 14, recipients were sacrificed, and implantation rates, estimated as the number of live fetuses plus resorptions, were determined. When ICSI was performed with sperm incubated in CM, no effects on fertilization, embryo cleavage, blastocyst rate, or blastocyst morphology were detected; however, the quality of the embryos was affected because the total implantation rate decreased significantly (P , .05) when 2-cell embryos or blastocysts were transferred. Independently of sperm pretreatment, in vitro cultures significantly affected the percentage of live fetuses present on day 14 of pregnancy. These results demonstrated that there are factors released from fragmented spermatozoa capable of inducing DNA fragmentation in intact sperm that may compromise, to some extent, birth rates after ICSI.

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“…Previous observations have suggested that long-term programming of postnatal development, growth and physiology can be irreversibly affected during the pre-implantation period of embryo development by suboptimal in vitro culture conditions. Several studies have reported a reduction in the post-implantation viability of embryos cultured in medium supplemented with serum [6][7][8][9]. It is thought that under these conditions the embryo does not receive the right cues, resulting in dysregulation of genes or aberrant epigenetic modifications in the genome [26].…”

Section: Discussionmentioning

confidence: 99%

“…The use of serum, a common component of culture media [3][4][5], provides a rich but undefined environment for embryonic development [3]. In fact, it has been found that serum reduces the post-implantation viability of embryos [6][7][8][9]. Previous results from our group have revealed that mice derived from embryos cultured in suboptimal conditions due to the addition of fetal calf serum (FCS) can develop obesity and display abnormally large hearts and livers in adulthood [6].…”

Section: Introductionmentioning

confidence: 99%

Hyperplastic Obesity and Liver Steatosis as Long-Term Consequences of Suboptimal In Vitro Culture of Mouse Embryos1

Serrano

Decara

Fernández‐González

et al. 2014

Biology of Reproduction

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In the present study, we identify and describe an obese phenotype in mice as a long-term consequence of a suboptimal in vitro culture that resulted from the addition of fetal calf serum (FCS) into the culture medium. Mice produced with FCS displayed a high mortality rate (approximately 55% versus 15% in control mice within 20 mo) and increased sensitivity to the development of obesity in adulthood when fed either a standard or a high-fat diet. These mice developed hyperplastic obesity that was characterized by a significant expansion of the fat pads (approximately 25% and 32% higher body weight in male and female mice over controls, respectively) with unchanged adipocyte size. We observed a sexual dimorphism in the development of obesity in the mice produced with FCS. Whereas the female mice displayed hypertension, hyperleptinemia, and fatty liver, the male mice only displayed glucose intolerance. The mRNA expression of metabolically relevant genes in the adipose tissue was also affected. The males produced with FCS expressed higher mRNA levels of the genes that activate fatty acid oxidation (peroxisome proliferator-activated receptor alpha [Ppara, PPARalpha] and acyl-CoA oxidase 1 [Acox1, ACOX1]) and thermogenesis (uncoupling protein 1 [Ucp1, UCP1]), which may counteract the metabolic phenotype. Conversely, the females produced with FCS generally expressed lower levels of these metabolic genes. In the females, the obese phenotype was associated with inhibition of the lipogenic pathway (peroxisome proliferator-activated receptor gamma [Pparg, PPARgamma] and fatty acid synthase [Fasn, FAS]), indicating a saturation of the storage capacity of the adipose tissue. Overall, our data indicate that the exposure to suboptimal in vitro culture conditions can lead to the sexually dimorphic development of obesity in adulthood.

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The effect of preimplantation culture conditions on murine embryo implantation and fetal development

Cited by 44 publications

References 20 publications

Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior

Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior

Factors From Damaged Sperm Affect Its DNA Integrity and Its Ability to Promote Embryo Implantation in Mice

Hyperplastic Obesity and Liver Steatosis as Long-Term Consequences of Suboptimal In Vitro Culture of Mouse Embryos1

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