Three Dimensional &amp;lt;i&amp;gt;In Vitro&amp;lt;/i&amp;gt; Culture of Murine Secondary Follicles in a Defined Synthetic Matrix

Asaduzzman, Mohammad; Cui, Xiaolin; Zhang, Hu; Young, Fiona

doi:10.4236/jbnb.2018.93014

Cited by 9 publications

(8 citation statements)

References 58 publications

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“…So, it seems that OCs can grow with the minimum essential materials but CCs need more constituents. Our results are parallel with some previous studies, that used DMEMF12 as the best media for OCs and CCs (14)(15). As well, in line with our results,…”

supporting

confidence: 93%

How to Support the In Vitro Culture of Human Ovarian Cells and Follicles? A Preliminary Step for Assembling an Artificial Ovary

Moshrefi

Aflatoonian

Ghasemi-Esmailabad

et al. 2021

Preprint

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BackgroundThe first step in assembling an artificial ovary is supporting the in vitro growth of ovarian cells/follicles. Therefore, proliferation of granulosa or cumulus cells (CCs) is important and due to limited proliferation, improving the mediums for in vitro propagation of these cells is helpful. ObjectiveThis study aimed to characterized the appropriate media and supplements for in vitro culture of ovarian cells (OCs) and CCs. Material & MethodsCortical, medullar and hilar cells of ovary were cultured and their conditioned medium (CMs) collected. The expression of GDF9, as a key factor for ovarian follicular growth, was evaluated. CCs were collected from healthy women, who referred due to male factor infertility. To choose the optimum basal medium, a mixture of OCs was cultured with basal mediums, supplemented with two concentrations of fetal bovine serum (FBS) and human serum albumin (HSA). The cocktails were as follows: [Serum free mediums], [mediums+10%FBS], [mediums+20%FBS], [mediums+1%Alb)], [mediums+2%Alb], [mediums+10%FBS+1%Alb], [mediums+10%FBS+2%Alb], [mediums+20%FBS+1%Alb] & [mediums+20%FBS+2%Alb]. The same process was repeated for CCs. Also, the effect of various concentrations of L-Glutamine, bovine serum albumin (BSA), HSA, insulin transferrin selenium (ITS), Follitropin alfa® and Pregnyl® was evaluated on the growth of CCs. As well, CCs were treated with various concentrations of follicular fluids (FFs) and CMs. CMs were collected from ovarian, testicular, adipose & amniotic derived and ovarian carcinoma cells. FFs were collected from women with poor responders, endometriosis, advanced age, polycystic ovarian syndrome, male factor and unknown infertility. Then, CCs morphology, proliferation capacity and culture duration were evaluated. ResultsAll the ovarian cells expressed GDF9. αMEM+20%FBS and DMEMF12+20%FBS were the most suitable cocktails for OCs and CCs, respectively. 20%FBS was superior to 10% for both OCs and CCs. HSA could not support the growth of OCs and CCs, alone. The cocktail of mediums with 20%FBS superior to the others. The CMs of cortical and (hillar+medullar) cells and FFs from male factor and unknown infertility patients caused higher CCs proliferation. 17 mM/l L-Glutamine, 24 mg/ml BSA, 20 mg/ml HSA, 10 ng/ml ITS, 300 mIU/ml Follitropinα and 3.5 IU/ml Pregnyl led to higher proliferation of CCs. ConclusionCMs, serums and FFs can support the CCs growth alongside with basal mediums, supplemented with hormones, ITS and L-Glutamine, which are cheaper and more accessible.

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supporting

confidence: 93%

How to Support the In Vitro Culture of Human Ovarian Cells and Follicles? A Preliminary Step for Assembling an Artificial Ovary

Moshrefi

Aflatoonian

Ghasemi-Esmailabad

et al. 2021

Preprint

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“…Murine secondary follicles were encapsulated in the 30 mg/ml SFX-504 1 and in vitro cultured for two days. Although the results demonstrated an increase in 505 follicles diameter from 153 ± 28 µm to 201 ± 38, only 12.5% of follicles maintained their 506 morphological integrity [196]. 507…”

Section: Other Natural and Synthetic Scaffolds 492mentioning

confidence: 88%

“…Moreover, the amount of estradiol and 500 progesterone produced by follicles seeded in gelatin/PCL was higher than PCL samples, 501 which indicated better follicle growth in this scaffold. SFX-1, a synthetic polymer, which was 502 produced from N-isopropylacrylamide (NIPAM) monomer, has also been used for ovarian 503 tissue engineering [196]. Murine secondary follicles were encapsulated in the 30 mg/ml SFX-504 1 and in vitro cultured for two days.…”

Section: Other Natural and Synthetic Scaffolds 492mentioning

confidence: 99%

A Review on Biomaterials for Ovarian Tissue Engineering

et al. 2021

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Considerable challenges in engineering the female reproductive tissue are the follicle's 25 unique architecture, the need to recapitulate the extracellular matrix, and tissue 26 vascularization. Over the years, various strategies have been developed for preserving 27 fertility in women diagnosed with cancer, such as embryo, oocyte, or ovarian tissue 28 cryopreservation. While autotransplantation of cryopreserved ovarian tissue is a viable 29 choice to restore fertility in prepubertal girls and women who need to begin chemo-or 30 radiotherapy soon after the cancer diagnosis, it is not suitable for all patients due to the risk 31 of having malignant cells present in the ovarian fragments in some types of cancer. Advances 32 in tissue engineering such as 3D printing and ovary-on-a-chip technologies have the 33 potential to be a translational strategy for precisely recapitulating normal tissue in terms of 34 physical structure, vascularization, and molecular and cellular spatial distribution. This 35 review first introduces the ovarian tissue structure, describes suitable properties of 36 biomaterials for ovarian tissue engineering, and highlights recent advances in tissue 37 engineering for developing an artificial ovary. 38 39 40 Keywords 41 Ovarian tissue engineering; follicle; in vitro culture; 3D printing; ovary-on-a-chip 42 157 9 Ovarian cells can be added to the isolated follicles to support their development 158 during in vitro culture or transplantation. These cells secrete autocrine/paracrine 159 factors involved in folliculogenesis [44], and some of them will be recruited by 160 growing follicles to differentiate into theca cells [48]. Moreover, to improve follicle 161 survival, one could play with the biomaterial used as a scaffold, its porosity, 162 mechanical strength, and bioactive sites, as well as the addition of hormones, growth 163 factors, etc. [8, 11, 49-53]. For instance, to enhance vascularization, follicles could be 164 encapsulated in biomaterials with sustained release of vascular endothelial growth 165 factor (VEGF) [54] or basic fibroblast growth factor (bFGF) [55]. 166 167 4 Development of follicle culture systems 168The 2D in vitro system has been routinely used for follicle' in vitro culture [56][57][58][59][60][61]. Several 169 approaches, such as multi-well plates, cell culture dishes, or coverslips coated by gels or ECM 170 proteins such as collagen, fibronectin, and laminin, have been used in this procedure [3, 33, 171 62]. When isolated follicles are attached to a 2D culture surface, the somatic cells migrate to 172 the bottom of the well, gradually losing their interaction with the oocyte, and consequently, 173 the follicle 3D structure is destroyed [7]. Indeed, 2D culture systems are different from in 174 vivo microenvironment conditions, where cells and tissues have complex connections with 709This study was supported by grants from the Fonds National de la Recherche Scientifique de 710 Belgique (the Excellence of Science (FNRS-EOS), number 30443682 (PhD scholarship 711 award...

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“…Follicle and oocyte diameters were assessed using a graticule eyepiece HWf10X-f DIN on 0, 3, 6, and 9 days of culture. 35 The percentage of follicles with eccentric oocytes was assessed on the mentioned days and antrum formation rate was recorded at the end of culture.…”

Section: Follicular and Oocyte Morphology And Viabilitymentioning

confidence: 99%

Co‐culture with peritoneum mesothelial stem cells supports the in vitro growth of mouse ovarian follicles

Sarabadani

Tavana

Mirzaeian

et al. 2021

J Biomedical Materials Res

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The important roles played by the ovarian microenvironment and cell interactions in folliculogenesis suggest promising approaches for in vivo growth of ovarian follicles using appropriate scaffolds containing suitable cell sources. In this study, we have investigated the growth of early preantral follicles in the presence of decellularized mesenteric peritoneal membrane (MPM), peritoneum mesothelial stem cells (PMSCs), and conditioned medium (CM) of PMSCs. MPM of mouse was first decellularized; PMSCs were isolated from MPM and cultured and their conditioned medium (CM) was collected. Mouse follicles were separated into four groups: (1) culture in base medium (control), (2) culture in decellularized MPM (DMPM), (3) co-culture with PMSCs (Co-PMSCs), and (4) culture in CM of PMSCs (CM-PMSCs). Qualitative and quantitative assessments were performed to evaluate intact mesenteric peritoneal membrane (IMPM) as well as decellularized ones. After culturing the ovarian follicles, follicular and oocyte diameter, viability, eccentric oocyte percentage, and estradiol hormone amounts were evaluated. Quantitative and qualitative evaluations confirmed removal of cells and retention of the essential fibers in MPM after the decellularization process. Follicular parameters showed that Co-PMSCs better support in vitro growth and development of ovarian follicles than the other groups. The eccentric rate and estradiol production were statistically higher for the Co-PMSCs group than for the CM-PMSCs and control groups. Although the culture of early preantral follicles on DMPM and CM-PMSCs could improve in vitro follicular growth, co-culture of follicles with PMSCs showed even greater improvements in terms of follicular growth and diameter. K E Y W O R D S conditioned medium, decellularized ECM, mesenteric peritoneal membrane, mesothelial stem cells, ovarian follicle culture | INTRODUCTIONChemo-and radiation therapies lead to early menopause, premature ovarian insufficiency (POI), and/or infertility in women. 1,2 One solution to this problem is the transplantation of fresh or frozen ovaries and ovarian tissues, but the risks of recurrence of malignancy in the cells and allogenic rejection of transplants have led to interest in the development of alternate techniques such as in vitro culture of isolated ovarian follicles. [2][3][4][5] Current follicular culture systems are inefficient due to inconsistent development of both somatic and germinal components of the ovarian follicles. 6 Moreover, in vitro cultured follicles have much smaller diameters and are poorer in quality than in vivo follicles. 7 There have been many studies in recent years aimed at improving in vitro follicle development through the use of various growth factors, supplements, and serum components. 6,8 The use of different matrices and co-cultures with supportive somatic cells and/or their

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Three Dimensional <i>In Vitro</i> Culture of Murine Secondary Follicles in a Defined Synthetic Matrix

Cited by 9 publications

References 58 publications

How to Support the In Vitro Culture of Human Ovarian Cells and Follicles? A Preliminary Step for Assembling an Artificial Ovary

How to Support the In Vitro Culture of Human Ovarian Cells and Follicles? A Preliminary Step for Assembling an Artificial Ovary

A Review on Biomaterials for Ovarian Tissue Engineering

Co‐culture with peritoneum mesothelial stem cells supports the in vitro growth of mouse ovarian follicles

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