Disposable set for cell culture, cell culture device and cell preparation method

A disposable set for cell culture comprising:

• (1) a cell culture container having a liquid inlet and a liquid outlet;
• (2) a cell separation kit for separating cells used for cell culture, the cell separation kit being connected to the liquid inlet,
• wherein the cell separation kit includes a cell separation material selectively trapping cells from a liquid to be treated containing the cells, the cell separation kit includes a cell separation filter storing the cell separation material,
• wherein the cell separation material is composed of fiber or fiber aggregate, the fiber or fiber aggregate has a diameter from 3 to 40 μm, the cell separation material has a density in a range from 1.0×104 to 1.0×106 g/m3, wherein the density is a value obtained by dividing the weight (g) of a cell separation material by the volume (m3), the cell separation material has an opening size of 3 μm or more in minor axis and an opening size of 120 μm or less in major axis, wherein the opening size is a mean value determined as follows:
• observing the cell separation material under a scanning electron microscope to give a micrograph;
• determining a major diameter and a minor diameter of a substantial hole formed by the intersections of two or more different fibers at fifty or more points with an image analyzer;
• and
• calculating each mean value calculated from the determined diameters;
• a tank for a liquid to be treated;
• wherein the tank stores the liquid to be treated containing cells,
• wherein the tank for the liquid to be treated is connected to an upstream side of the cell separation filter via a first pipeline,
• a drain tank connected to a downstream side of the cell separation filter via a second pipeline,
• a first cell collection liquid introduction part for introducing a cell collection liquid,
• wherein the first cell collection liquid introduction part is a syringe storing the cell collection liquid and a plunger of the syringe is pressurized by an external pressure unit driven by gas pressure to supply the cell collection liquid to the cell separation filter,
• a piston rod operably connected to the plunger, the plunger having a plurality of positions including a position A and a position B, the piston rod having a plurality of positions including a first position and second position, wherein the piston rod is configured to move from the first position to the second position from the external pressure thereby moving the plunger from position A to position B and thereby pushing out the cell collection liquid and causing the cell collection liquid to flow to the cell separation filter, and wherein the plunger and the piston rod are configured to return to position A and the first position, respectively, through the external pressure unit driven by gas pressure,
• the first cell collection liquid introduction part being connected by a pipeline branched at a midway point of the second pipeline, and a third pipeline branched at a midway point of the first pipeline and connected to the liquid inlet of the cell culture container;
• a cell storage bag for once storing cells collected by the cell collection liquid introduced from the first cell collection liquid introduction part to the cell separation filter, wherein the cell storage bag is connected to a pipeline branched at a more upstream position of the first pipeline relative to the third pipeline, wherein the once storing cells is a time of collection from the cell separation filter;
• and
• (3) a cell collection kit for washing and concentrating cells cultured in the cell culture container, the cell collection kit being connected to the liquid outlet,
• wherein the tank for a liquid to be treated is configured as the cell storage bag once storing cells collected by the cell collection liquid introduced from the first cell collection liquid introduction part to the cell separation filter,
• wherein the cell separation filter substantially traps cells useful for cell medicine or regenerative medicine by sending a liquid to be treated containing the cells useful for cell medicine or regenerative medicine and impurity cells to substantially pass the impurity cells through the cell separation filter, and the trapped cells are collected by passing the cell collection liquid in a direction opposite to the direction in which the liquid to be treated is passed, and
• wherein a washing liquid tank storing a washing liquid is connected to an upstream side of the cell separation filter via a pipeline.