Evaluation of a new freezing protocol containing 20% dimethyl sulphoxide concentration to cryopreserve human ovarian tissue

Gallardo, Miguel; Paulini, Fernanda; Corral, Ariadna; Balcerzyk, Marcin; Lucci, Carolina Madeira; Ambroise, Jérôme; Merola, Marta; Fernández-Maza, Laura; Risco, Ramón; Dolmans, Marie-Madeleine; Amorim, Christiani Andrade

doi:10.1016/j.rbmo.2018.09.012

Cited by 23 publications

(14 citation statements)

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“…The use of human ovaries was approved by the Institutional Review Board of the Université Catholique de Louvain on May 2019 (IRB reference 2012/23MAR/125). The ovaries were frozen, based on the Gallardo et al protocol [ 53 ]. After tissue thawing, stromal cells were isolated, as previously described by Asiabi et al [ 54 ].…”

Section: Methodsmentioning

confidence: 99%

Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

Jafari

Dadashzadeh

Moghassemi

et al. 2021

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An artificial ovary is a promising approach for preserving fertility in prepubertal girls and women who cannot undergo current cryopreservation strategies. However, this approach is in its infancy, due to the possible challenges of creating a suitable 3D matrix for encapsulating ovarian follicles and stromal cells. To maintain the ovarian stromal cell viability and proliferation, as a first step towards developing an artificial ovary, in this study, a double network hydrogel with a high water swelling capacity (swelling index 15–19) was developed, based on phenol conjugated chitosan (Cs-Ph) and silk fibroin (SF) through an enzymatic crosslinking method using horseradish peroxidase. The addition of SF (1%) to Cs (1%) decreased the storage modulus (G’) from 3500 Pa (Cs1) to 1600 Pa (Cs-SF1), and the hydrogels with a rapid gelation kinetic produced a spatially homogeneous distribution of ovarian cells that demonstrated 167% proliferation after 7 days. This new Cs-SF hydrogel benefits from the toughness and flexibility of SF, and phenolic chemistry could provide the potential microstructure for encapsulating human ovarian stromal cells.

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Section: Methodsmentioning

confidence: 99%

Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

Jafari

Dadashzadeh

Moghassemi

et al. 2021

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“…The ovarian tissue biopsies were collected from postmenopausal women (51-68 years old) and transferred to the laboratory in Dulbecco's phosphate-buffered saline (DPBS; 14,190,144;Gibco). The ovarian cortical fragments were frozen using our standard method after the medullar section was removed [14]. After thawing of the ovarian samples, stromal cells were isolated following our published protocols [12,[15][16][17].…”

Section: Ovarian Stromal Cell Isolationmentioning

confidence: 99%

Medium supplementation can influence the human ovarian cells in vitro

et al. 2022

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Background Cells are an essential part of the triple principles of tissue engineering and a crucial component of the engineered ovary as they can induce angiogenesis, synthesize extracellular matrix and influence follicle development. Here, we hypothesize that by changing the medium supplementation, we can obtain different cell populations isolated from the human ovary to use in the engineered ovary. To this end, we have in vitro cultured cells isolated from the menopausal ovarian cortex using different additives: KnockOut serum replacement (KO), fetal bovine serum (FBS), human serum albumin (HSA), and platelet lysate (PL). Results Our results showed that most cells soon after isolation (pre-culture, control) and cells in KO and FBS groups were CD31- CD34- (D0: vs. CD31-CD34+, CD31 + CD34+, and CD31 + CD34- p < 0.0001; KO: vs. CD31-CD34+, CD31 + CD34+, and CD31 + CD34- p < 0.0001; FBS: vs. CD31-CD34+ and CD31 + CD34+ p < 0.001, and vs. CD31 + CD34- p < 0.01). Moreover, a deeper analysis of the CD31-CD34- population demonstrated a significant augmentation (more than 86%) of the CD73+ and CD90+ cells (possibly fibroblasts, mesenchymal stem cells, or pericytes) in KO- and FBS-based media compared to the control (around 16%; p < 0.001). Still, in the CD31-CD34- population, we found a higher proportion (60%) of CD90+ and PDPN+ cells (fibroblast-like cells) compared to the control (around 7%; vs PL and KO p < 0.01 and vs FBS p < 0.001). Additionally, around 70% of cells in KO- and FBS-based media were positive for CD105 and CD146, which may indicate an increase in the number of pericytes in these media compared to a low percentage (4%) in the control group (vs KO and FBS p < 0.001). On the other hand, we remarked a significant decrease of CD31- CD34+ cells after in vitro culture using all different medium additives (HSA vs D0 p < 0.001, PL, KO, and FBS vs D0 P < 0.01). We also observed a significant increase in epithelial cells (CD326+) when the medium was supplemented with KO (vs D0 p < 0.05). Interestingly, HSA and PL showed more lymphatic endothelial cells compared to other groups (CD31 + CD34+: HSA and PL vs KO and FBS p < 0.05; CD31 + CD34 + CD90 + PDPN+: HSA and PL vs D0 p < 0.01). Conclusion Our results demonstrate that medium additives can influence the cell populations, which serve as building blocks for the engineered tissue. Therefore, according to the final application, different media can be used in vitro to favor different cell types, which will be incorporated into a functional matrix.

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“…玻璃化冻存的另一个瓶颈是其在较大组织器官中应用时的去玻璃化问题. 组织、器官成分复杂, 细胞类型、胞间成分多样, 极易造成 CPA 分布不均匀以及局部冷却速率不均而出现温度梯度, 进而出现胞间连接断裂, 这些断裂间隙的产生可能提供更多的冰成核位点, 增加升温复苏过程中去玻璃化发生的风险 [87] . 此外, 由于组织和器官的组成复杂性, 各部分的热导率和热容不均一, 所以很难在复温过程中实现均匀传热, 解冻过程的冰晶重结晶对组织造成的损伤也不容忽视 [88][89] .…”

Section: 玻璃化冻存unclassified

Bio-inspired Ice-controlling Materials for Cryopreservation of Cells and Tissues

Zheng¹,

Liu²,

Liu³

et al. 2021

Acta Chimica Sinica

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Evaluation of a new freezing protocol containing 20% dimethyl sulphoxide concentration to cryopreserve human ovarian tissue

Cited by 23 publications

References 43 publications

Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

Medium supplementation can influence the human ovarian cells in vitro

Bio-inspired Ice-controlling Materials for Cryopreservation of Cells and Tissues

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