Composition of protein supplements used for human embryo culture

Morbeck, Dean E.; Paczkowski, Melissa; Fredrickson, J.R.; Krisher, Rebecca L.; H, Hoff; Baumann, N; Moyer, Thomas P.; Matern, Dietrich

doi:10.1007/s10815-014-0349-2

Cited by 69 publications

(37 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Primers for genes previously correlated with embryo viability [6,32,33] (bone morphogenic protein 15, Bmp15; caudaltype homeobox protein 2, Cdx2; cyclooxgenase 2, Cox2; DNA methyltransferase 3a, Dnmt3a; glutaredoxin2, Glrx2; glucose transporter 1, Glut1; octamer-binding protein 4, Oct4), and the reference gene peptidylprolylisomerase A (Ppia) were designed for quantitative PCR (qPCR) using Primer3 [31], and qPCR was performed as previously described [6,32,33]. Accession number, primer sequence, and product length of target and reference genes are described in Supplemental Table 1.…”

Section: Quantitative Pcrmentioning

confidence: 99%

“…product should be used for clinical ART. The more troubling scenario is the possibility that a product could support the development of murine embryos but still be unsuitable for clinical use [3,6]. The key to preventing such products from entering the clinic is to improve the sensitivity of MEAs and ensure clinically relevant results.…”

Section: Introductionmentioning

confidence: 99%

“…There are multiple ways to alter the sensitivity of a MEA, including changing the developmental stage of the embryo at the initiation of culture, the conditions used for culture, or the strain of mouse [4,[6][7][8]. All of these manipulations effectively alter the amount of stress placed on the embryo during culture.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Herrick

Paik

Strauss

et al. 2015

J Assist Reprod Genet

Self Cite

View full text Add to dashboard Cite

Purpose The aim of this study is to compare the sensitivity of the standard one-cell mouse embryo assay (MEA) to that using in vitro-matured oocytes from hybrid and outbred mice. Methods The study was done by culturing embryos in the presence or absence of two concentrations (0.0005 or 0.001 %v/v) of Triton X-100 (TX100). Embryonic development, blastocyst cell numbers (total and allocation to the trophectoderm [TE] and inner cell mass [ICM]), and blastocyst gene expression were evaluated. Results Neither concentration of TX100 affected (P>0.05) cleavage, blastocyst development, or hatching in one-cell embryos from BDF1 mice. However, all cell number endpoints were reduced (P<0.05) by the high concentration of TX100 and the number of ICM cells was reduced (P<0.05) by the low concentration of TX100. Inhibitory (P<0.05) effects of the high concentration of TX100 were observed in in vitro maturation (IVM) embryos from BDF1, CF1, and SW, but not ICR, mice. Cell number and allocation were negatively affected by the high concentration of TX100 in CF1 and SW embryos, but not in BDF1 or ICR embryos. The only developmental endpoints affected by the low concentration of TX100 were cleavage of BDF1 oocytes, blastocyst development of SW embryos, and cell numbers (total and inner cell mass (ICM)) of SW blastocysts. Conclusions The sensitivity of the MEA to TX100 is improved by using embryos from in vitro-matured oocytes, using oocytes from some outbred (SW or CF1, not ICR) strains of mice, and evaluating blastocyst cell number and allocation.

show abstract

Section: Quantitative Pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Herrick

Paik

Strauss

et al. 2015

J Assist Reprod Genet

Self Cite

View full text Add to dashboard Cite

show abstract

“…Previous studies have demonstrated that both HSA and complex protein supplements are heterogeneous in nature [19] and thus represent variation that may impact MEA results. Specifically, complex proteins (SPS, LGPS) contain more transition metals than other protein supplements which may increase reactive-oxidant species and result in cell damage [18]. This study found SPS increased toxicity while HSA did not affect sensitivity.…”

Section: Discussionmentioning

confidence: 78%

“…For human IVF, decreased oxygen concentration (5%) is both physiologic in the reproductive tract and associated with improved outcomes [13][14][15][16], yet many IVF clinics throughout the world still use atmospheric oxygen for culture and for QC testing [17]. The type of protein also varies in practice, thus, representing another variable that could influence assay sensitivity [18,19]. A third variation in embryo culture methods, group versus individual culture, influences QC sensitivity [10], and evidence suggests that group culture improves IVF outcomes, either through diffusion of unknown paracrine factors or maintenance of a stable microenvironment [20].…”

Section: Introductionmentioning

confidence: 99%

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Ainsworth

Fredrickson

Morbeck

2017

J Assist Reprod Genet

Self Cite

View full text Add to dashboard Cite

Purpose Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. Methods Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations. Assessment of blastocyst development at either 96 h or in an extended MEA (eMEA) to 144 h was tested in each experiment. In experiment 1, ability to detect peroxides in oil was compared in the MEA, eMEA, and cell number at 96 h. In experiment 2, serial dilutions of peroxide in oil were used along with time-lapse imaging to compare sensitivity of the morphokinetic MEA to the eMEA. Culture conditions that may affect assay sensitivity were assessed in experiments 3-5, which examined the effect of group versus individual culture, oxygen concentration, and protein supplementation. Results Extended MEA and cell counts identified toxicity not detected by the routine endpoint of blastocyst rate at 96 h. The eMEA was fourfold more sensitive than the standard MEA, and this sensitivity was similar to the morphokinetic MEA. Group culture had a protective effect against toxicity, while oxygen concentration did not affect blastocyst development. Protein supplementation with HSA had a protective effect on blastocyst development in eMEA.Conclusions The standard MEA used by manufacturers does not detect potentially lethal toxicity of peroxides in mineral oil. While group culture may mask toxicity, protein supplementation and oxygen concentration have minimal effect on assay sensitivity. The eMEA and time-lapse morphokinetic assessment are equally effective in detection of peroxide toxicity and thus provide manufacturers and end-users a simple process modification that can be readily adopted into an existing QC program.

show abstract

The Female Reproductive Tract and Early Embryo Development

Leese¹,

Brison²

2018

Clinical Reproductive Science

View full text Add to dashboard Cite

Composition of protein supplements used for human embryo culture

Cited by 69 publications

References 46 publications

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Improved detection of mineral oil toxicity using an extended mouse embryo assay

The Female Reproductive Tract and Early Embryo Development

Contact Info

Product

Resources

About