The present disclosure provides a method for preparing heterogeneous hematopoietic stem and progenitor cells using non-mobilized peripheral blood, which uses a capsule culture system to capture and proliferate rare hematopoietic stem and progenitor cells in non-mobilized peripheral blood, and prepares heterogeneous hematopoietic stem and progenitor cell clones. The present disclosure captures the rare heterogeneous stem cells in non-mobilized peripheral blood and morphologically verifies the presence of heterogeneous hematopoietic stem and progenitor cells in non-mobilized peripheral blood. The method of the present disclosure has the characteristics of hematopoietic reconstitution, drug development, transplantation and immunotherapy, gene editing of cell types, and the like. The method of the present disclosure provides a reliable cell source for patient-specific functional hematopoietic stem cells, and actively promotes the clinical application of non-mobilized hematopoietic stem and progenitor cells.