scholarly journals MiR-191-5p Is Upregulated in Culture Media of Implanted Human Embryo on Day Three of Development

2020 ◽  
Author(s):  
Ricardo Josue Acuña-González ◽  
Fela Vanesa Morales-Hernández ◽  
Jorge Skiold López-Canales ◽  
Jair Lozano-Cuenca ◽  
Mauricio Osorio-Caballero ◽  
...  
Keyword(s):  
Embryo Development ◽  
Human Embryo ◽  
Culture Medium ◽  
Culture Media ◽  
Expression Profiles ◽  
Uterine Cavity ◽  
Common Criteria ◽  
Significant Difference ◽  
Human Embryo Development

Abstract Background: Morphologic features are the most common criteria for selecting human embryo to be transferred to the receptive uterine cavity. However, such characteristics are not valid for embryos in cellular arrest. The aim of this study was to quantify the expression profile of hsa-miR-21-3p, -24-1-5p, -191-5p, and -372-5p on day 3 of culture media from in vitro fertilization (IVF) embryo that were implanted or failed to be implanted in patients (n=25 pregnant and 25, non-pregnant patients). Methods: Fifty patients were accepted in the Department of Reproductive Biology of a Hospital in México City, based on the Institutional inclusion criteria for in vitro fertilization. On day 3 of development, embryos were transferred to women, and the culture medium was collected from implanted embryos (n=25, pregnant patients) and non-implanted embryos (n=25, non-pregnant). In the culture medium, RNA was isolated using TRIzol reagent. MiRNA expression was detected through RT-PCR with specific primers. Expression bands were quantified using an optic density.Results: The expression profiles were compared between pregnant and non-pregnant patients revealing a significant 5.2-fold greater expression of hsa-miR-191-5p in the former group (p ≤0.001) and a significantly higher expression of hsa-miR-24-1-5p (p =0.043) in the latter. No significant difference was found between the two groups in regard hsa-miR-21-3p or hsa-miR-372-5p (p =0.41). Conclusions: According to the results, has-miR-191-5p could possibly be a possible biomarker of adequate human embryo development. This miRNA modulated IGF2BP-1 and IGF2R, which are associated with the implantation window. On the other hand, hsa-miR-24-1-5p may be related to a poor prognosis of human embryo development.

scholarly journals MiR-191-5p is upregulated in culture media of implanted human embryo on day fifth of development

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Ricardo Josué Acuña-González ◽  
Mercedes Olvera-Valencia ◽  
Jorge Skiold López-Canales ◽  
Jair Lozano-Cuenca ◽  
Mauricio Osorio-Caballero ◽  
...  
Keyword(s):  
Embryo Development ◽  
Human Embryo ◽  
Poor Prognosis ◽  
Culture Media ◽  
Uterine Cavity ◽  
Human Embryos ◽  
Common Criteria ◽  
Significant Difference ◽  
The Media

Abstract Background Morphological features are the most common criteria used to select human embryos for transfer to a receptive uterine cavity. However, such characteristics are not valid for embryos in cellular arrest. Even aneuploid embryos can have normal morphology, and some euploid embryos have aberrant morphology. The aim of this study was to quantify the expression profile of hsa-miR-21-3p, -24-1-5p, -191-5p, and -372-5p in culture media on day 5 of in vitro embryo development, and compare the profiles of two groups of media classified by outcome: successful (n = 25) or unsuccessful (n = 25) implantation pregnancy. Methods Fifty patients were accepted in the Department of Reproductive Biology of a Hospital in México City, based on the Institutional inclusion criteria for in vitro fertilization. Embryos were transferred to the women on day 5 of cultivation, and the culture media were collected. RNA was isolated from each culture medium with TRIzol reagent, and microRNA (miRNA) expression was detected through RT-PCR with specific primers. Expression bands were quantified by reading optical density. Results There was a 5.2-fold greater expression of hsa-miR-191-5p in the pregnancy-related culture media (p ≤ 0.001) and a 1.6-fold greater level of hsa-miR-24-1-5p (p = 0.043) in the media corresponding to non-pregnant women. No significant difference existed between the two groups hsa-miR-21-3p (p = 0.38) or hsa-miR-372-5p (p = 0.41). Conclusions Regarding adequate in vitro embryo development, hsa-miR-191-5p could possibly represent a positive biomarker, while has-miR-24-1-5p may indicate poor prognosis. This former miRNA modulates IGF2BP-1 and IGF2R, associated with the implantation window. On the other hand, hsa-miR-24-1-5p may be related to a poor prognosis of human embryo development.

scholarly journals Integrating insulin into single-step culture medium regulates human embryo development in vitro

2017 ◽  
Vol 107 (2) ◽  
pp. 405-412 ◽  
Author(s):  
Mohamed Fawzy ◽  
Mohamed Sabry ◽  
Mohamed Nour ◽  
Mohamed Y. Abdelrahman ◽  
Eman Roshdy ◽  
...  
Keyword(s):  
Embryo Development ◽  
Human Embryo ◽  
Culture Medium ◽  
Single Step ◽  
Human Embryo Development ◽  
Development In Vitro

160 COMPARATIVE STUDY OF IN VITRO CULTURE MEDIA AND ASSISTED HATCHING TECHNIQUES ON MOUSE EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 188
Author(s):  
N. C. Negota ◽  
L. P. Nethenzheni ◽  
M. L. Mphaphathi ◽  
D. M. Barry ◽  
T. L. Nedambale
Keyword(s):  
In Vitro Culture ◽  
Chorionic Gonadotropin ◽  
Culture Medium ◽  
Culture Media ◽  
Assisted Hatching ◽  
Hatching Rate ◽  
Significant Difference ◽  
Equine Chorionic Gonadotropin ◽  
F1 Generation

The in vitro culture media and assisted hatching techniques remain challenging obstacles to be utilised widely. Mechanical, chemical, enzymatic thinning, and laser-assisted techniques have been used previously but information is still lacking on its application in livestock. The aim of this study was to compare the effect of 2 in vitro culture media (Hamster F10 and TMC-199) and 4 (mechanical, chemical, enzymatic, and laser) assisted hatching techniques on blastocyst formation and hatching rate using murine embryos as a model. The C57/b and Balb/c breeds were raised until they reached maturity and bred naturally to produce F1 generation. The light in the breeding house was controlled at 14 h light and 10 h dark. Feed and water were provided ad libitum for the mice. Superovulation of females were stimulated using equine chorionic gonadotropin and human chorionic gonadotropin. The F1 generation was used for the collection of the 400 blastocysts and randomly allocated into 4 assisted hatching techniques. Blastocysts were paired into a group of 10 and replicated 4 times for each assisted hatching technique. The general linear model of SAS version 9.4 (SAS Institute Inc., Cary, NC, USA) was used to analyse the data. Assisted hatching techniques of laser, mechanical, enzymatic, and chemical yielded 46.9 ± 37.1, 51.1 ± 40.2, 39.1 ± 35.8, and 33.3 ± 4.5%, respectively, under in vitro culture of Hamster F10. The TCM-199, laser, mechanical, enzymatic, and chemical assisted hatching techniques yielded 56.3 ± 43.3, 52.6 ± 35.5, 49.2 ± 37.5, and 33.9 ± 35.5%, respectively, with a significant difference. There was no significant difference observed in assisted hatching techniques and Hamster F10 culture medium. However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM than cultured in Hamster F10. Hatching rate of blastocysts increased from chemical, enzymatic, mechanical, and laser with response to Hamster F10 and TCM; thus, laser is a suitable assisted hatching technique with TCM-199.

Impairment of Human Embryo Development after Abnormal in Vitro Fertilization

1985 ◽  
Vol 442 (1 In Vitro Fert) ◽  
pp. 336-341 ◽  
Author(s):  
MICHELLE PLACHOT ◽  
JACQUELINE MANDELBAUM ◽  
ANNE-MARIE JUNCA ◽  
JACQUES SALAT-BAROUX ◽  
JEAN COHEN
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Human Embryo ◽  
Vitro Fertilization ◽  
Human Embryo Development

scholarly journals Embryo Development of Cypripedium formosanum in Relation to Seed Germination In Vitro

2005 ◽  
Vol 130 (5) ◽  
pp. 747-753 ◽  
Author(s):  
Yung-I Lee ◽  
Nean Lee ◽  
Edward C. Yeung ◽  
Mei-Chu Chung
Keyword(s):  
Seed Germination ◽  
Embryo Development ◽  
Developmental Stages ◽  
Culture Media ◽  
Nile Red ◽  
Germination Percentage ◽  
Zygotic Embryogenesis ◽  
Seed Collection ◽  
Significant Difference

This investigation documents the key anatomical features in embryo development of Cypripedium formosanum Hayata, in association with the ability of embryos to germinate in vitro, and examines the effects of culture media and seed pretreatments on seed germination. A better understanding of zygotic embryogenesis for the Cypripedium L. species would provide insights into subsequent germination events and aid in the in vitro propagation of these endangered species. In seeds collected at 60 days after pollination (DAP), soon after fertilization, no germination was recorded. The best overall germination was found at 90 DAP (≈70%), at which time early globular to globular embryos with a single-celled suspensors can be observed. After 135 DAP, the seeds germinated poorly. At this time the inner integument shrinks and forms a tight layer, which encloses the embryo, the so-called “carapace.” Using Nile red stain, a cuticular substance was detected in the carapace, which may play a role in the impermeability of the mature seed and may help the seeds survive in the stringent environment. At maturity (after 210 DAP), the embryo proper has an average size of eight cells along its length and six cells across the width. Lipids and proteins are the main storage products within the embryo. To improve seed germination, experiments were conducted to test the suitability of various media and pretreatments of seeds. When different media were used, except for the Harvais medium at 120 DAP, there was no significant difference in seed germination at three different developmental stages tested. Soaking mature seeds in 1% NaOCl or treating them with ultrasound may slightly increase the germination percentage. For seed germination, our results indicate that the timing of seed collection outweighs the composition of medium and the seed pretreatments.

scholarly journals Cell-Free, Embryo-Specific sncRNA as a Molecular Biological Bridge between Patient Fertility and IVF Efficiency

2019 ◽  
Vol 20 (12) ◽  
pp. 2912 ◽  
Author(s):  
Angelika V. Timofeeva ◽  
Vitaliy V. Chagovets ◽  
Yulia S. Drapkina ◽  
Nataliya P. Makarova ◽  
Elena A. Kalinina ◽  
...  
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Expression Profiles ◽  
Embryo Implantation ◽  
Human Reproduction ◽  
Vitro Fertilization ◽  
Small Noncoding Rnas ◽  
Polymerase Chain ◽  
Next Generation Sequencing Ngs

Small noncoding RNAs (sncRNAs) are key regulators of the majority of human reproduction events. Understanding their function in the context of gametogenesis and embryogenesis will allow insight into the possible causes of in vitro fertilization (IVF) implantation failure. The aim of this study was to analyze the sncRNA expression profile of the spent culture media on day 4 after fertilization and to reveal a relationship with the morphofunctional characteristics of gametes and resultant embryos, in particular, with the embryo development and implantation potential. Thereto, cell-free, embryo-specific sncRNAs were identified by next generation sequencing (NGS) and quantified by reverse transcription coupled with polymerase chain reaction (RT-PCR) in real-time. Significant differences in the expression level of let-7b-5p, let-7i-5p, piR020401, piR16735, piR19675, piR20326, and piR17716 were revealed between embryo groups of various morphological gradings. Statistically significant correlations were found between the expression profiles of piR16735 and piR020401 with the oocyte-cumulus complex number, let-7b-5p and piR020401 with metaphase II oocyte and two pronuclei embryo numbers, let-7i-5p and piR20497 with the spermatozoid count per milliliter of ejaculate, piR19675 with the percentage of linearly motile spermatozoids, let-7b-5p with the embryo development grade, and let-7i-5p with embryo implantation. According to partial least squares discriminant analysis (PLS-DA), the expression levels of let-7i-5p (Variable Importance in Projection score (VIP) = 1.6262), piR020401 (VIP = 1.45281), and piR20497 (VIP = 1.42765) have the strongest influences on the implantation outcome.

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