Hyaluronic acid-alginate hydrogel stimulates the differentiation of neonatal mouse testicular cells into hepatocyte-like and other cell lineages in three-dimensional culture

IF 2.4 4区生物学 Q4 CELL BIOLOGY

Biology of the Cell Pub Date : 2024-08-23 DOI:10.1111/boc.202400049

Leila Rashki Ghaleno, Mohammad Amin Hajari, Mahmoud Alipour Choshali, Elham Abed Heidari, Abdolhossein Shahverdi, Hiva Alipour, Mojtaba Rezazadeh Valojerdi

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Abstract

Background information

Extracellular matrix (ECM)-derived hydrogels are frequently used in three-dimensional (3D) cell culture and organoid formation in several tissues. However, in the 3D cultivation of testicular cells, the hyaluronic acid (HA) hydrogel has not received as much attention. This study examined the effects of three distinct composites, including HA-alginate (HA-Alg), HA-alginate-collagen (HA-Alg-Col), and HA-alginate-decellularized ECM (HA-Alg-dECM), on mouse testicular cell culture and in vitro spermatogenesis.

Methods

For the creation of composites, the concentration of biomaterials used was 0.5% HA, 1% alginate, 2.5 mg/mL collagen, and 25 mg/mL dECM derived from the testicles of Rams. After 3D culture of 5 days post-partum (dpp) mouse testicular cells for 14 days, HA-Alg was selected as a superior composite due to the greater number and size of the produced organoids. Then, cell culture was rerun by HA-Alg for 14 days, which was later extended for an additional 28 days. In addition, the 3D culture of 10 dpp mouse testicular cells was used to compare with 5 dpp mice on day 14. The morphology and gene expression were analyzed using appropriate techniques.

Results

On day 14, the HA-Alg hydrogel showed significantly more organoids in terms of size and number than the other two groups (p < 0.05); nevertheless, none of the groups showed the expected signs of testis organoids. Remarkably, on day 14, the histology and immunostaining tests revealed features of hepatocyte-like cells (HLCs) and albumin production as a marker of HLC functionality. Furthermore, the analysis of gene expression verified the significant expression of angiogenesis markers (p < 0.01). After the extended culture to 28 days, 5 dpp testicular cells once more differentiated into erythrocytes and HLCs, while a small number of organoids showed the characteristic of renal cells. Cell culture of 10 dpp mice for 14 days showed a wide range of cell lineages, including renal, glandular, chondrocyte, and hepatocyte-like cells in comparison to the 5 dpp mice.

Conclusion and significance

While the HA-Alg composite did not support spermatogenesis in the 3D culture of mouse testicular cells, it demonstrated an unpredicted potential for promoting the differentiation of neonate mouse testicular cells into HLC, erythrocytes, and other cell lineages.

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透明质酸-精氨酸水凝胶刺激新生小鼠睾丸细胞在三维培养中分化为肝细胞样和其他细胞系。

背景信息：细胞外基质（ECM）衍生的水凝胶常用于多种组织的三维（3D）细胞培养和类器官形成。然而，在睾丸细胞的三维培养中，透明质酸（HA）水凝胶并没有受到如此多的关注。本研究考察了三种不同的复合材料，包括 HA-海藻酸盐（HA-Alg）、HA-海藻酸盐-胶原蛋白（HA-Alg-Col）和 HA-海藻酸盐-去细胞化 ECM（HA-Alg-dECM）对小鼠睾丸细胞培养和体外精子发生的影响：在制作复合材料时，使用的生物材料浓度为 0.5% HA、1% 藻酸盐、2.5 mg/mL 胶原和 25 mg/mL 来自公羊睾丸的 dECM。在对产后 5 天（dpp）的小鼠睾丸细胞进行 14 天的三维培养后，HA-Alg 被选为一种更优的复合材料，因为其产生的器官组织数量更多，体积更大。然后，用 HA-Alg 重新进行细胞培养 14 天，之后又延长了 28 天。此外，10 dpp 小鼠睾丸细胞的三维培养在第 14 天与 5 dpp 小鼠进行了比较。结果：结果：第 14 天，HA-Alg 水凝胶组的器官大小和数量明显多于其他两组（p 结论和显著性：在小鼠睾丸细胞的三维培养过程中，HA-Alg 复合材料并不支持精子发生，但它在促进新生小鼠睾丸细胞分化为 HLC、红细胞和其他细胞系方面表现出了意想不到的潜力。

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来源期刊

Biology of the Cell 生物-细胞生物学

CiteScore

5.30

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： The journal publishes original research articles and reviews on all aspects of cellular, molecular and structural biology, developmental biology, cell physiology and evolution. It will publish articles or reviews contributing to the understanding of the elementary biochemical and biophysical principles of live matter organization from the molecular, cellular and tissues scales and organisms. This includes contributions directed towards understanding biochemical and biophysical mechanisms, structure-function relationships with respect to basic cell and tissue functions, development, development/evolution relationship, morphogenesis, stem cell biology, cell biology of disease, plant cell biology, as well as contributions directed toward understanding integrated processes at the organelles, cell and tissue levels. Contributions using approaches such as high resolution imaging, live imaging, quantitative cell biology and integrated biology; as well as those using innovative genetic and epigenetic technologies, ex-vivo tissue engineering, cellular, tissue and integrated functional analysis, and quantitative biology and modeling to demonstrate original biological principles are encouraged.