Ra Kihae, Oh Hyun Ju, Kim Eun Young, Kang Sung Keun, Ra Jeong Chan, Kim Eui Hyun, Park Se Chang, Lee Byeong Chun
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea.
Research and Development Center, MKbiotech Co., Ltd., 99 Daehak-ro, Daejeon 34134, Korea.
Antioxidants (Basel). 2021 Feb 9;10(2):268. doi: 10.3390/antiox10020268.
Oxidative stress is a major cause of damage to the quantity and quality of embryos produced in vitro. Antioxidants are usually supplemented to protect embryos from the suboptimal in vitro culture (IVC) environment. Amniotic membrane-derived mesenchymal stem cells (AMSC) have emerged as a promising regenerative therapy, and their paracrine factors with anti-oxidative effects are present in AMSC conditioned medium (CM). We examined the anti-oxidative potential of human AMSC-CM treatment during IVC on mouse preimplantation embryo development and antioxidant gene expression in the forkhead box O (FoxO) pathway. AMSC-CM (10%) was optimal for overall preimplantation embryo developmental processes and upregulated the expression of FoxOs and their downstream antioxidants in blastocysts (BL). Subsequently, compared to adipose-derived mesenchymal stem cell (ASC)-CM, AMSC-CM enhanced antioxidant gene expression and intracellular GSH levels in the BL. Total antioxidant capacity and SOD activity were greater in AMSC-CM than in ASC-CM. Furthermore, SOD and catalase were more active in culture medium supplemented with AMSC-CM than in ASC-CM. Lastly, the anti-apoptotic effect of AMSC-CM was observed with the regulation of apoptosis-related genes and mitochondrial membrane potential in BL. In conclusion, the present study established AMSC-CM treatment at an optimal concentration as a novel antioxidant intervention for assisted reproduction.
氧化应激是体外产生的胚胎数量和质量受损的主要原因。通常会添加抗氧化剂来保护胚胎免受体外培养(IVC)环境欠佳的影响。羊膜来源的间充质干细胞(AMSC)已成为一种有前景的再生疗法,其具有抗氧化作用的旁分泌因子存在于AMSC条件培养基(CM)中。我们研究了在IVC期间人AMSC-CM处理对小鼠植入前胚胎发育以及叉头框O(FoxO)途径中抗氧化基因表达的抗氧化潜力。AMSC-CM(10%)对植入前胚胎的整体发育过程最为适宜,并上调了囊胚(BL)中FoxO及其下游抗氧化剂的表达。随后,与脂肪来源的间充质干细胞(ASC)-CM相比,AMSC-CM增强了BL中的抗氧化基因表达和细胞内谷胱甘肽水平。AMSC-CM中的总抗氧化能力和超氧化物歧化酶(SOD)活性高于ASC-CM。此外,添加AMSC-CM的培养基中SOD和过氧化氢酶的活性高于ASC-CM。最后,通过调节BL中凋亡相关基因和线粒体膜电位观察到了AMSC-CM的抗凋亡作用。总之,本研究确定了最佳浓度的AMSC-CM处理作为辅助生殖的一种新型抗氧化干预措施。