Improving in vitro induction efficiency of human primordial germ cell-like cells using N2B27 or NAC-based medium
Gege Yuan1
Jiachen Wang1
Shuangshuang Qiu1
Yunfei Zhu1
Qing Cheng2
Laihua Li1
Jiahao Sha1
Xiaoyu Yang3
Yan Yuan1
1.State Key Laboratory of Reproductive Medicine and Offspring Health,Nanjing Medical University,Nanjing,Jiangsu 211166,China2.Women's Hospital of Nanjing Medical University,Women and Children's Healthcare Hospital,Nanjing,Jiangsu 211100,China3.State Key Laboratory of Reproductive Medicine,Clinical Center of Reproductive Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210029,China
摘要：Primordial germ cells(PGCs),the precursors of oocytes and spermatozoa,are highly pluripotent.In recent years,the in vitro induction of human primordial germ cell-like cells(hPGCLCs)has advanced significantly.However,the stability and efficacy of obtaining hPGCLCs in vitro still require improvement.In the current study,we identified a novel induction system using Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12)as the basal medium,supplemented with B27 and N2(referred to as N2B27)in combination with four cytokines:bone morphogenetic protein 4,stem cell factor,epidermal growth factor,and leukemia inhibitory factor.The hPGCLCs induced under these conditions closely resembled PGCs from 4-to 5-week-old embryos at the transcriptomic level.Compared with traditional GK15(GMEM supplemented with 15％KnockoutTM Serum Replacement)-based induction conditions,the N2B27 system significantly increased the speed and efficacy of hPGCLC induction.RNA sequencing analysis revealed that this improvement was due to an increased cellular capacity to cope with hypoxic stress and avoid apoptosis.The N2B27 medium promoted enhanced mitochondrial activity,enabling cells to better manage hypoxic stress while reducing the production of reactive oxygen species.Moreover,through gradient concentration experiments,we demonstrated that the addition of the common antioxidant N-acetyl-L-cysteine at an optimized concentration further enhanced the efficiency of PGCLC induction under GK15 conditions.In summary,we have established an optimized induction system that enhances the efficiency of hPGCLC differentiation by improving cellular resilience to hypoxic stress and apoptosis.
机标关键词：vitrohumancellsgermcell-likeefficiencyimprovinginduction
分类号：R321.1(人体形态学)
论文发表日期：2025-11-30
在线出版日期：2026-01-15（本平台首次上网日期，不代表文献的发表时间）
页数：15( 587-600,中插11-中插13 )
英文信息