Francés-Herrero Emilio, De Miguel-Gómez Lucía, López-Martínez Sara, Campo Hannes, Garcia-Dominguez Ximo, Diretto Gianfranco, Faus Amparo, Vicente José S, Marco-Jiménez Francisco, Cervelló Irene
Fundación Instituto Valenciano de Infertilidad (FIVI), Instituto de Investigación Sanitaria La Fe, Avenida Fernando Abril Martorell, 106. Hospital La Fe, Torre A, Planta 1ª, 46026, Valencia, Spain.
Universitat de València, Valencia, Spain.
Reprod Sci. 2021 Jun;28(6):1644-1658. doi: 10.1007/s43032-020-00446-6. Epub 2021 Jan 28.
The oviducts (fallopian tubes in mammals) function as the site of fertilization and provide necessary support for early embryonic development, mainly via embryonic exposure to the tubal microenvironment. The main objective of this study was to create an oviduct-specific extracellular matrix (oviECM) hydrogel rich in bioactive components that mimics the native environment, thus optimizing the developmental trajectories of cultured embryos. Rabbit oviducts were decellularized through SDS treatment and enzymatic digestion, and the acellular tissue was converted into oviductal pre-gel extracellular matrix (ECM) solutions. Incubation of these solutions at 37 °C resulted in stable hydrogels with a fibrous structure based on scanning electron microscopy. Histological staining, DNA quantification and colorimetric assays confirmed that the decellularized tissue and hydrogels contained no cellular or nuclear components but retained important components of the ECM, e.g. hyaluronic acid, glycoproteins and collagens. To evaluate the ability of oviECM hydrogels to maintain early embryonic development, two-cell rabbit embryos were cultured on oviECM-coated surfaces and compared to those cultured with standard techniques. Embryo development was similar in both conditions, with 95.9% and 98% of the embryos reaching the late morula/early blastocyst stage by 48 h under standard culture and oviECM conditions, respectively. Metabolomic analysis of culture media in the presence or absence of embryos, however, revealed that the oviECM coating may include signalling molecules and release compounds beneficial to embryo metabolism.
输卵管(哺乳动物的输卵管)作为受精场所,主要通过胚胎暴露于输卵管微环境为早期胚胎发育提供必要支持。本研究的主要目的是制备一种富含生物活性成分的输卵管特异性细胞外基质(oviECM)水凝胶,模拟天然环境,从而优化培养胚胎的发育轨迹。通过SDS处理和酶消化使兔输卵管脱细胞,将脱细胞组织转化为输卵管前凝胶细胞外基质(ECM)溶液。基于扫描电子显微镜,这些溶液在37℃孵育后形成具有纤维结构的稳定水凝胶。组织学染色、DNA定量和比色分析证实,脱细胞组织和水凝胶不含细胞或核成分,但保留了ECM的重要成分,如透明质酸、糖蛋白和胶原蛋白。为了评估oviECM水凝胶维持早期胚胎发育的能力,将二细胞兔胚胎培养在涂有oviECM的表面,并与采用标准技术培养的胚胎进行比较。在两种条件下胚胎发育相似,在标准培养和oviECM条件下,分别有95.9%和98%的胚胎在48小时时达到桑葚胚晚期/囊胚早期阶段。然而,对有无胚胎存在时培养基的代谢组学分析表明,oviECM涂层可能包含信号分子并释放对胚胎代谢有益的化合物。