National Clinical Research Center for Obstetrical and Gynecological Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Key Laboratory of Cancer Invasion and Metastasis, Ministry of Education, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Climacteric. 2022 Apr;25(2):170-178. doi: 10.1080/13697137.2021.1921726. Epub 2021 May 17.
The aim of this study was to design and fabricate a three-dimensional (3D) printed artificial ovary.
We first compared the printability of gelatin-methacryloyl (GelMA), alginate and GelMA-alginate bioinks, of which GelMA was selected for further investigation. The swelling properties, degradation kinetics and shape fidelity of GelMA scaffolds were characterized by equilibrium swelling/lyophilization, collagenase processing and micro-computed tomography evaluation. Commercial ovarian tumor cell lines (COV434, KGN, ID8) and primary culture ovarian somatic cells were utilized to perform cell-laden 3D printing, and the results were evaluated by live/dead assays and TUNEL detection. Murine ovarian follicles were seeded in the ovarian scaffold and their diameters were recorded every day. Finally, maturation was performed, and the ovulated oocytes were collected and observed.
Our results indicated that GelMA was suitable for 3D printing fabrication. Its scaffolds performed well in terms of hygroscopicity, degradation kinetics and shape fidelity. The viability of ovarian somatic cells was lower than that of commercial cell lines, suggesting that extrusion-based 3D culture fabrication is not suitable for primary ovarian cells. Nevertheless, the GelMA-based 3D printing system provided an appropriate microenvironment for ovarian follicles, which successfully grew and ovulated in the scaffolds. Metaphase II oocytes were also observed after maturation.
The GelMA-based 3D printing culture system is a viable alternative option for follicular growth, development and transfer. Accordingly, it shows promise for clinical application in the treatment of female endocrine and reproductive conditions.
本研究旨在设计并制造一种三维（3D）打印人工卵巢。
我们首先比较了明胶甲基丙烯酰（GelMA）、海藻酸盐和 GelMA-海藻酸盐生物墨水的可打印性，选择 GelMA 用于进一步研究。通过平衡溶胀/冷冻干燥、胶原酶处理和微计算机断层扫描评估来表征 GelMA 支架的溶胀性能、降解动力学和形状保真度。利用商业卵巢肿瘤细胞系（COV434、KGN、ID8）和原代培养的卵巢体细胞进行细胞负载的 3D 打印，并通过活/死检测和 TUNEL 检测评估结果。将小鼠卵巢卵泡接种在卵巢支架中，并每天记录其直径。最后，进行成熟培养，并收集和观察排卵的卵母细胞。
我们的结果表明 GelMA 适合 3D 打印制造。其支架在吸湿性、降解动力学和形状保真度方面表现良好。卵巢体细胞的活力低于商业细胞系，这表明基于挤出的 3D 培养制造不适用于原代卵巢细胞。然而，基于 GelMA 的 3D 打印系统为卵巢卵泡提供了适宜的微环境，卵泡在支架中成功生长和排卵。在成熟培养后，也观察到了中期 II 期卵母细胞。
基于 GelMA 的 3D 打印培养系统是卵泡生长、发育和转移的可行替代方案。因此，它有望在治疗女性内分泌和生殖疾病的临床应用中得到应用。
