The invention discloses a totally-enclosed cell and cell factor preparation device which comprises an upper blood bag, a lower blood bag and a thin connecting pipe, wherein the upper blood bag is provided with an anti-coagulation inlet, a plasma sucking port for sucking plasma and an output port for outputting cells and cell factors; the bottom of the upper blood bag is conical; the top of the lower blood bag is conical; and the upper blood bag, the thin connecting pipe and the lower blood bag are formed into a dumbbell-shaped blood collecting bag. The cell and cell factor preparation method comprises the following steps of: introducing anti-coagulation blood, marrow and cord blood into the blood collecting bag, erecting the blood collecting bag on a hard sleeve, putting the hard sleeve into a centrifuge, and starting the centrifuge; separating the lower blood bag from the thin connecting pipe; fully mixing the plasma in the upper blood bag, and extracting the plasma in the upper blood bag for testing platelet concentration; reserving the plasma in the upper blood bag for later use; or centrifuging again to extract a plasma layer, and reserving centrifugally precipitated cells. The invention has the advantages of simplicity and convenience in operation, low loss of platelets, mononuclear cells and cell factors and high enrichment ratio, and obviously improves the safety and practicality.