Protective effects of trehalose on frozen-thawed ovarian granulosa cells of cattle

Zheng, Yuxin; Ma, Lizhu; Liu, S.J.; Zhang, C.T.; Meng, Rui; Chen, Y.Z.; Jiang, Zhongliang

doi:10.1016/j.anireprosci.2018.11.005

Cited by 8 publications

(3 citation statements)

References 31 publications

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“…It is at concentrations higher than 0.2 mol/L [48]. The overexpression of α-SMA is a pathological hallmark of the activation of HSCs and their transition to myofibroblasts [49].…”

Section: Journal Of Nanomaterialsmentioning

confidence: 99%

Preparation and Characterization of PEG-PLA Genistein Micelles Using a Modified Emulsion-Evaporation Method

Cheng

Qin

et al. 2020

Journal of Nanomaterials

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The objective of this study is to improve the bioavailability of genistein by encapsulation with polyethylene glycol-polylactic acid (PEG-PLA) copolymers. Genistein micelles (GMs) prepared using a modified emulsion-evaporation method were more stable than those made with the original method. The effect of polyvinyl alcohol, Tween 80, sonication time, PEG-PLA/genistein ratio, and organic phase (acetone)/H2O ratio on the size, polydispersity index, encapsulation efficiency, and drug loading efficiency of GMs was investigated. GMs were obtained and characterized under optimal experimental conditions. For long-term storage, GMs were lyophilized by freeze drying with trehalose to produce genistein lyophilized powder (GLP). The analysis of GLP by Fourier-transform infrared spectroscopy and differential scanning calorimetry showed that genistein was successfully incorporated into the micellar structure. In vitro release experiments revealed that the incorporation of genistein into PEG-PLA copolymers significantly improved its solubility and bioavailability. GLP was more potent in inhibiting the proliferation of HSC-T6 cells than genistein. Treatment with GLP at 10–20 μg/mL for 48 h significantly inhibited the protein expression of α-smooth muscle actin and collagen I in HSC-T6 cells compared with the control. These data demonstrated that the improved solubility and bioavailability of genistein in the form of GLP enhanced its antifibrotic effect in vitro.

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“…It is at concentrations higher than 0.2 mol/L [48]. The overexpression of α-SMA is a pathological hallmark of the activation of HSCs and their transition to myofibroblasts [49].…”

Section: Journal Of Nanomaterialsmentioning

confidence: 99%

Preparation and Characterization of PEG-PLA Genistein Micelles Using a Modified Emulsion-Evaporation Method

Cheng

Qin

et al. 2020

Journal of Nanomaterials

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“…The cells at a density of 8 × 10 5 /mL cells per well were treated with 0.5 μM U73122 or m3M3FBS for 4 h. The concentration of intracellular Ca 2+ was monitored by a fluorescent Ca 2+ indicator Fluo 3-AM (Sigma, St. Louis, MO, USA), and was measured as described previously [23]. The intracellular Ca 2+ changes were measured with the ratio of fluorescence intensities excited at 488 nm and 530 nm using flow cytometry (CyFlow Cube, PARTEC, Germany).…”

Section: Methodsmentioning

confidence: 99%

Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

et al. 2019

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Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 mRNA expression was decreased after treatment with m-3M3FBS (the PLC activator). Both the early and late apoptosis rate were maximized with 0.5 μM U73122 for 4 h. The rate of early apoptosis was the highest at 4 h and the rate of late apoptosis was the highest at 12 h in the m-3M3FBS group. The protein abundance of PLCβ1, protein kinase C β (PKCβ), calmodulin-dependent protein kinaseII α (CAMKIIα) and calcineurinA (CalnA) were decreased by U73122, and CAMKIIα protein abundance was increased by m-3M3FBS. The mRNA expression of several downstream genes (CDC42, NFATc1, and NFκB) was upregulated by PLC. Our results demonstrated that apoptosis can be inhibited by altering PLC signaling in porcine primary granulosa cells cultured in vitro, and several calcium−sensitive targets and several downstream genes might take part in the processes.

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“…There are several studies demonstrating cryopreservation of ovarian tissues and cells in various species, including human and bovine (Amorim et al 2011;Baufeld and Vanselow 2018;Bouillon et al 2020;Kokotsaki et al 2018;Pietrowski et al 2020;Rivas Leonel et al 2019;Santana et al 2012;Shi et al 2017;Sluss et al 1994;Youm et al 2014;Zheng et al 2019). They indicate that freezing/thawing is, in general, possible, yet the survival rate of cells is rather low (45-58%; see Bouillon et al 2020).…”

Section: Introductionmentioning

confidence: 99%

A rapid and robust method for the cryopreservation of human granulosa cells

Beschta

Eubler²,

Bohne

et al. 2021

Histochem Cell Biol

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Human primary granulosa cells (GCs) derived from women undergoing oocyte retrieval can be cultured and used as a cellular model for the study of human ovarian function. In vitro, they change rapidly, initially resembling cells of the preovulatory follicle and then cells of the corpus luteum. They are derived from individual patients, whose different medical history, lifestyle and age lead to heterogeneity. Thus, cells can rarely be ideally matched for cellular experiments or, if available, only in small quantities. We reasoned that cryopreservation of human GCs may be helpful to improve this situation. Previous studies indicated the feasibility of such an approach, but low survival of human GCs was reported, and effects on human GC functionality were only partially evaluated. We tested a slow freezing protocol (employing FCS and DMSO) for human GCs upon isolation from follicular fluid. We compared cryopreserved and subsequently thawed cells with fresh, non-cryopreserved cells from the same patients. About 80% of human GCs survived freezing/thawing. No differences were found in cell morphology, survival rate in culture, or transcript levels of mitochondrial (COX4, OPA1, TOMM20), steroidogenic (CYP11A1, CYP19A1) or cell–cell contact genes (GJA1) between the two groups in cells cultured for 1–5 days. A proteomic analysis revealed no statistically significant change in the abundance of a total of 5962 proteins. The two groups produced comparable basal levels of progesterone and responded similarly to hCG with elevation of progesterone. Taken together, our results show this to be a rapid and readily available method for the cryopreservation of human GCs. We anticipate that it will allow future large-scale experiments and may thereby improve cellular studies with human ovarian cells.

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Protective effects of trehalose on frozen-thawed ovarian granulosa cells of cattle

Cited by 8 publications

References 31 publications

Preparation and Characterization of PEG-PLA Genistein Micelles Using a Modified Emulsion-Evaporation Method

Preparation and Characterization of PEG-PLA Genistein Micelles Using a Modified Emulsion-Evaporation Method

Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

A rapid and robust method for the cryopreservation of human granulosa cells

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