Herein, human POI-iPSCs with genotype of POI, including fragile X syndrome, abnormal karyotype (45, X; 45, X/46, XX; 45, XO and 47, XXX), and the gene mutation (FIGLA and GDF9), were generated using a lentiviral vector with four reprogramming factors under the 4i culture system. Here, GDF9 is linked to fragile X syndrome.