The invention provides a composition for amplifying hematopoietic stem cells and a culture medium containing the same, a kit, a pharmaceutical composition, an application and a method for amplifying the hematopoietic stem cells. The composition is made of JNK-IN-8, nicotinamide adenine dinucleotide, rapamycin and Y27632. The composition of the invention can efficiently amplify the hematopoietic stem cells, effectively maintain protein phenotype and gene expression profile of the hematopoietic stem cells, improve in vivo reconstruction function and reconstruction efficiency of the hematopoieticstem cells and has great scientific research, clinical research and application values.