In vitro maturation on an agarose matrix improves the developmental competence of porcine oocytes

Park, Ji Eun; Kim, Minji; Lee, Joohyeong; Chai, Changhoon; Lee, Seung Tae; Lee, Eunsong

doi:10.1016/j.theriogenology.2020.07.016

Cited by 8 publications

(7 citation statements)

References 55 publications

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“…Based on the presence of integrin subunits α 2 , α 5 , α 6 , β 1 , and β 4 on the surface of CCs, fibronectin interacting with integrin α 5 β 1 , collagen interacting with integrin α 2 β 1 , and laminin interacting with integrin α 6 β 1 or α 6 β 4 were incorporated into 1% (w/v) agarose matrices. In this study, 1% (w/v) agarose gels were applied because they display similar mechanical properties to ovarian cortices (Park et al., 2020). Immature COCs derived from porcine MAFs were matured in vitro on each acellular niche, which consisted of different combinations of ECM proteins incorporated into IVM agarose matrices.…”

Section: Discussionmentioning

confidence: 99%

“…However, these factors are not represented in current IVM systems. In a previous study, we attempted to develop an acellular niche based on the physical properties of the ovarian cortex by producing an agarose matrix that mimicked ovarian cortex stiffness (herein referred to as IVM agarose matrix) (Park et al., 2020). However, the mechanical factors of our matrix were insufficient to stimulate cytoplasmic maturation in oocytes in vitro , which suggested that additional stimulators should be incorporated into the agarose matrix to more accurately reflect the mechanics of ovarian cortices.…”

Section: Introductionmentioning

confidence: 99%

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In vitro maturation using an agarose matrix with incorporated extracellular matrix proteins improves porcine oocyte developmental competence by enhancing cytoplasmic maturation

Park

Kim

Lee

et al. 2021

J Tissue Eng Regen Med

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Here, we present a novel in vitro maturation (IVM) system comprising an agarose matrix supplemented with extracellular matrix (ECM) proteins for enhanced maturation of immature oocytes within cumulus–oocyte complexes (COCs) derived from porcine medium antral follicles (MAFs). Immunocytochemical analyses of integrin subunit α2, α5, α6, β1, and β4 expression suggested that integrin α2β1, α5β1, α6β1, and α6β4 play pivotal roles in IVM of porcine immature oocytes. Combinatorial supplementation of fibronectin interacting with integrin α5β1, collagen interacting with integrin α2β1, and laminin interacting with integrin α6β1 and α6β4 to the agarose matrix had no significant effect on nuclear maturation. However, the number of parthenogenetic embryos that developed into blastocysts increased when oocytes were matured using agarose IVM matrices supplemented with fibronectin, collagen, or laminin. Furthermore, significant increases in cytoplasmic maturation‐related parameters (BMP15 level, cumulus cell expansion score, intra‐oocyte ATP level, and index of cortical granule distribution) were observed in COCs matured in vitro using ECM protein‐incorporated agarose matrices. Our data suggest that mature porcine oocytes with enhanced developmental competence and high‐quality cytoplasm can be generated via IVM using agarose matrices supplemented with fibronectin, collagen, or laminin.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro maturation using an agarose matrix with incorporated extracellular matrix proteins improves porcine oocyte developmental competence by enhancing cytoplasmic maturation

Park

Kim

Lee

et al. 2021

J Tissue Eng Regen Med

Self Cite

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“…The primer sequences used in qPCR were designed with Primer Premier 6.0 (Premier Biosoft International) software (Table 1). The relative quantification of gene expression was analysed using the 2 −ΔΔCt method (Park et al., 2020). All operations of relative quantifications complied with the Minimal Information for Publication of Quantitative Real‐Time PCR Experiments (MIQE) guidelines (Bustin et al., 2009).…”

Section: Methodsmentioning

confidence: 99%

Low oxygen concentration improves yak oocyte maturation and inhibits apoptosis through HIF‐1 and VEGF

Zhang

et al. 2022

Reprod Domestic Animals

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The gas-phase environment of in vitro culture system plays an important role in the development of oocytes, and oxygen concentration is one of the important factors. In the present study, we aimed to explore the effect of different oxygen concentrations (20%, 10%, 5% or 1% O 2 ) in yak oocyte maturation and to detect the expression of hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF) and cell apoptosis in yak COCs. First, the maturation rate of oocytes, cleavage rate and blastocysts rate following parthenogenetic activation in the group with 5% oxygen concentration were significantly higher (p < .05) than the other groups. Then, TUNEL analysis showed that the 5% oxygen concentration group significantly inhibited apoptosis of cumulus-oocyte complexes (COCs) compared to the other group, and the transcription and protein levels of pro-apoptotic factor Bax, HIF-1α and VEGF in yak COCs significantly reduced, while anti-apoptotic factor Bcl-2 significantly increased.Furthermore, immunohistochemical staining results indicated that HIF-1α protein was mainly located in theca follicle interna, mural follicular stratum granulosum, corona radiata and ovarian stroma in the follicular ovarian tissue, while VEGF protein was mainly located in the granulosa and theca cell layers. In summary, our findings demonstrate that 5% oxygen concentration may promote maturation and inhibit apoptosis of oocytes through HIF-1α-mediated VEGF expression.

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“…[28] Previous studies have reported a decrease in the proportion of TUNEL-positive cells across various cell types following 3D hydrogel culture. [11,29,30] However, this study represents the first investigation of DNA integrity, specifically in oocytes. We found that the integrity of DNA in oocytes from the bilayer hydrogel system was similar to both controls (2D and Alg; p > 0.05) and that DNA fragmentation rates did not exceed 15% (Figure 5B).…”

Section: Chromatin Configurationmentioning

confidence: 99%

“…However, there is limited research on 3D cell culture systems during IVM, and, in the majority of these studies, hydrogels are used chiefly as scaffolds to maintain the 3D structure of cells during culture and have rather obviate properties, such as hydrogel stiffness. [9][10][11][12][13] Tissues are characterized by varying degrees of stiffness. [14] In the context of reproductive tissues, the microenvironment in which the oocytes are growing is a combination of tissues exhibiting markedly different levels of stiffness.…”

Section: Introductionmentioning

confidence: 99%

A Biomimetic Follicle‐Based Design for Engineering Reproductive Technologies

Sánchez‐Ajofrín,

Andreu,

Galindo

et al. 2023

Adv Funct Materials

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In the field of assisted reproductive technologies, oocyte in vitro maturation (IVM) has emerged as a potential alternative to overcome the limitations associated with standard ovarian stimulation treatments. However, current IVM protocols lack standardization and yield oocytes of inferior quality compared to those matured in vivo. To address this issue, novel biomaterials like hydrogels offer distinct advantages in cell culture by providing a 3D cellular environment and facilitating easy adjustment and characterization of mechanical properties, such as hydrogel stiffness. Herein, a novel and reusable bilayer hydrogel system is presented that accurately mimics the mechanical properties of the microenvironment surrounding oocyte maturation. The system comprises an outer layer fabricated from either a 3D‐printed synthetic polymer (2‐vinyl‐4,6‐diamino‐1,3,5‐triazine, (VDT)) or a natural polymer (chitosan), along with an inner layer composed of alginate. By faithfully replicating the mechanical properties of native tissue within a 3D culture environment, this system significantly improves the quality and developmental capacity of oocytes, thus resulting in successful embryo development. Overall, this innovative system holds great potential for future medical research and applications in cell culture, thus representing a significant advancement in assisted reproductive technologies.

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In vitro maturation on an agarose matrix improves the developmental competence of porcine oocytes

Cited by 8 publications

References 55 publications

In vitro maturation using an agarose matrix with incorporated extracellular matrix proteins improves porcine oocyte developmental competence by enhancing cytoplasmic maturation

In vitro maturation using an agarose matrix with incorporated extracellular matrix proteins improves porcine oocyte developmental competence by enhancing cytoplasmic maturation

Low oxygen concentration improves yak oocyte maturation and inhibits apoptosis through HIF‐1 and VEGF

A Biomimetic Follicle‐Based Design for Engineering Reproductive Technologies

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