In vitro culture of mechanically isolated murine primary follicles in the presence of human platelet lysate PLTMax.

IF 1.8 Q3 OBSTETRICS & GYNECOLOGY Jornal Brasileiro de Reproducao Assistida Pub Date : 2024-08-26 DOI:10.5935/1518-0557.20240008

Vera Lucia Lângaro Amaral, Jhuly Laurentino Nunes, Rafael Alonso Salvador, Alfred Paul Senn, Tiago Góss Dos Santos

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Abstract

Objective: To develop a system for the culture of murine preantral ovarian follicles using Human Serum Albumin (HSA) and Human Platelet Lysate (PLTMax).

Methods: Mechanically isolated preantral follicles (N=146) were obtained from Swiss mice and cultured in DMEM:F12 medium for ten days in a 96-well plate with conical bottom. The medium was supplemented with penicillin, streptomycin, and equine chorionic gonadotropin. Additional proteins were tested in 4 test groups: G1: human serum albumin (HSA), G2: human platelet lysate (PLTM), and G3 and G4: HSA + PLTMax at lower and higher concentrations, respectively. Cellular vitality and oocyte morphology were evaluated on day 11 of culture.

Results: The highest follicular growth (3.4 fold) was achieved in HSA (G1), while a significantly lower (1.8 fold) growth was achieved in the presence of PLTM (G2, G4) and even further reduced (1.2 fold) when HSA and PLTM were combined (G3). Cellular vitality was close to 70-80% among the four groups, and the highest number of intact oocytes were found in G1.

Conclusions: PLTM did not improve follicular development and oocyte maturation compared to HSA but preserved cell vitality.

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在人血小板裂解液 PLTMax 的存在下对机械分离的小鼠原始卵泡进行体外培养。

目的：利用人血清白蛋白（HSA）和人血小板裂解液（PLTMax）开发小鼠前卵巢卵泡培养系统：开发一种使用人血清白蛋白（HSA）和人血小板裂解液（PLTMax）培养小鼠前卵巢卵泡的系统：方法：从瑞士小鼠身上获取机械分离的前卵巢卵泡（N=146），在 DMEM:F12 培养基中培养 10 天，培养基为 96 孔锥形底板。培养基中添加了青霉素、链霉素和马绒毛膜促性腺激素。在 4 个试验组中测试了其他蛋白质：G1：人血清白蛋白（HSA）；G2：人血小板裂解物（PLTM）；G3 和 G4：HSA + PLTMax，浓度分别较低和较高。在培养的第 11 天对细胞活力和卵母细胞形态进行评估：结果：HSA（G1）的卵泡生长率最高（3.4 倍），而 PLTM（G2、G4）的卵泡生长率明显较低（1.8 倍），当 HSA 和 PLTM 结合使用（G3）时，卵泡生长率甚至进一步降低（1.2 倍）。四组的细胞活力接近 70-80%，G1 中完整卵母细胞的数量最多：与 HSA 相比，PLTM 没有改善卵泡发育和卵母细胞成熟，但保持了细胞活力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Jornal Brasileiro de Reproducao Assistida OBSTETRICS & GYNECOLOGY-

CiteScore

3.30

自引率

6.70%

发文量