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成功地在生物工程化 Wharton 胶凝胶中水培培养和移植了小鼠分离的原始腔前卵泡。

Successful 3D culture and transplantation of mouse isolated preantral follicles in hydrogel of bioengineered Wharton's jelly.

机构信息

Department of Embryology, Royan Institute for Reproductive Biomedicine, Reproductive Biomedicine Research Center, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran.

Faculty of Veterinary Medicine, Garmsar Branch, Islamic Azad University, Garmsar, Iran.

出版信息

PLoS One. 2023 Sep 20;18(9):e0290095. doi: 10.1371/journal.pone.0290095. eCollection 2023.

DOI:10.1371/journal.pone.0290095
PMID:37729236
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10511077/
Abstract

MAIN OBJECTIVE

Due to Human Wharton's Jelly (HWJ) could be applied in tissue engineering as a bio scaffold, the present study was conducted to investigate the effects of HWJ hydrogel on in vitro culture and auto-transplantation of mouse ovarian follicles.

MATERIALS AND METHODS

HWJ was isolated from umbilical cord and decellularized with SDS/Tris/EDTA. DNA, Collagen and Glycosaminoglycans (GAGs) were measured. Decellularized Wharton's Jelly (DWJ) was dissolved to make Wharton's Jelly Hydrogel (WJH), and composited with Alginate (ALG) (1.5%) in equal ratio (WJH+ALG). Then, mouse preantral follicles were isolated and encapsulated in 10μL droplets of WJH and randomly considered for both 14 days culture and auto-transplantation.

RESULTS

Collagen, GAGs and DNA evaluations showed majority of WJ cells have been removed and MTT approved no toxicity. Degradation rate and rheological analysis represented optimal hydrogel compatibility. The data from in vitro culture revealed significant antral formation in WJH+ALG (P≤0.05). In transplantation, follicles failed to survive in ALG; however, survived in WJH+ALG to antral stage (P<0.05). VEGF and CD34 had greater expression in WJH+ALG than ALG (P< 0.05).

CONCLUSION

Wharton's jelly hydrogel and Alginate compound is interesting composite for successful development of mouse preantral follicles in both 3D in vitro culture and transplantation.

摘要

目的

由于人脐带华通氏胶（HWJ）可作为生物支架应用于组织工程，本研究旨在探讨 HWJ 水凝胶对体外培养和小鼠卵巢卵泡自体移植的影响。

材料和方法

HWJ 从脐带中分离出来，并用 SDS/Tris/EDTA 脱细胞化。测量 DNA、胶原和糖胺聚糖（GAGs）。将脱细胞化的华通氏胶（DWJ）溶解制成华通氏胶水凝胶（WJH），并与藻酸盐（ALG）（1.5%）以相等的比例（WJH+ALG）复合。然后，分离出小鼠原始卵泡并包裹在 10μL 的 WJH 液滴中，随机考虑进行 14 天培养和自体移植。

结果

胶原、GAGs 和 DNA 评估表明，WJ 细胞的大部分已被去除，MTT 证实无毒性。降解率和流变学分析表明最佳的水凝胶相容性。体外培养数据显示 WJH+ALG 组中出现明显的腔前形成（P≤0.05）。在移植中，卵泡在 ALG 中无法存活；然而，在 WJH+ALG 中存活并发育到腔前阶段（P<0.05）。WJH+ALG 中的 VEGF 和 CD34 表达高于 ALG（P<0.05）。

结论

华通氏胶水凝胶和藻酸盐复合物是成功开发小鼠原始卵泡的 3D 体外培养和移植的有趣组合。

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