The invention provides a method for preparing autologous hematopoietic stem cells. The method includes following steps of cultivating somatic cells in a cell culture fluid for 3-6 days and collecting the generated autologous hematopoietic stem cells. The cell culture fluid is an RPMI1640 culture fluid containing: 1-100 [mu]M of Y-27632 (C14H21N3O.2HCl), 1-100 ng/ml of stem cell factors, 1-100 ng/ml of interleukin 3, 1-100 ng/ml of interleukin 6, 1-100 ng/ml of interleukin 11, 1-100 ng/ml of a macrophage colony stimulating factor (M-CSF), 1-100 ng/ml of a granulocyte colony-stimulating factor (G-CSF), 1-100 [mu]g/ml of fucoidin, 1-100 [mu]g/ml of dextran sulfate, 1-100 nM of AMD3100 (1,1'-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetrazacyclotetradecane), 1-100 [mu]M of M-alendronate sodium (Malendronate sodium trihydrate), and 1-100 [mu]M of disodium pamidronate. The invention provides a method for preparing the autologous hematopoietic stem cells, a kit, the stem cells and an application. In addition, the method is advantaged in yield, purity, and producing speed of the production of the protein-induced autologous hematopoietic stem cells.