Stem cell separating material and method of separation
View Patent ↗The present invention has its object to provide a material for separating stem cell and a filter for separating stem cell, each is capable of selectively separating and recovering, in a simple and easy manner, stem cells from body fluids or biological tissue-derived treated fluids, a method for separating and recovering stem cells, and stem cells obtained by such method. The present invention is a material for separating stem cell which has a density K of 1.0×10 4 ≦K≦1.0×10 6 and a fiber diameter of 3 to 40 μm; a filter for separating stem cell which comprises the material for separating stem cell as packed in a container having a fluid inlet port and a fluid outlet port; a method of separating and recovering stem cells which comprises using the material for separating stem cell or the filter for separating stem cell; and a method of producing a multipotent cell fraction.
1. A method of separating and recovering stem cells from a body fluid or a biological tissue-derived treated fluid
which comprises
introducing a body fluid or the biological tissue-derived treated fluid into a filter comprising a material for separating stem cell having a density K of 1.0×10 4 ≦K≦1.0×10 6 g/m 3 and a fiber diameter of 3 to 40 μm, as packed in a container having a fluid inlet port and a fluid outlet port, through the fluid inlet port thereof, and
a cell recovering fluid is caused to flow from the fluid outlet side to thereby recover adherent stem cells captured by the material for separating stem cell,
and further culturing the adherent stem cells in a culture dish, a culture flask, or a culture bag,
wherein the adherent stem cells are mesenchymal stem cells or bone marrow stromal cells,
wherein the body fluid is bone marrow fluid, and
wherein the biological tissue-derived treated fluid is a treated fluid obtained by disintegration of at least one biological tissue selected from the group consisting of subcutaneous fat, visceral fat, white fat, and brown fat.
2. The method of separating and recovering stem cells according to claim 1 ,
wherein
a body fluid or biological tissue-derived treated fluid is introduced into a filter comprising a material for separating stem cell having a density K of 1.0×10 4 ≦K≦1.0×10 6 g/m 3 and a fiber diameter of 3 to 40 μm, as packed in a container having a fluid inlet port and a fluid outlet port, through the fluid inlet port thereof,
a washing fluid is caused to flow from the fluid inlet side for washing and then
a cell recovering fluid is caused to flow from the fluid outlet side to thereby recover adherent stem cells captured by the material for separating stem cell.
3. The method of separating and recovering stem cells according to claim 1 ,
wherein the body fluid or biological tissue is of the mammalian origin.
4. The method of separating and recovering stem cells according to claim 1 ,
wherein a minor axis of each mesh opening is not shorter than 3 μm and a major axis thereof is not longer than 120 μm.
5. The method of separating and recovering stem cells according to claim 4 ,
wherein the material is made of at least one synthetic polymer selected from among polyesters, rayon, polyolefins, vinylon, polystyrene, acrylics, nylons and polyurethanes.
6. The method of separating and recovering stem cells according to claim 4 ,
wherein the material is made of a combination of synthetic polymers, namely a polyester and polypropylene; rayon and a polyolefin; or a polyester, rayon and vinylon.
7. The method of separating and recovering stem cells according to claim 4 ,
wherein the material for separating stem cell substantially allows the passage of erythrocytes and leukocytes therethrough.
8. The method of separating and recovering stem cells according to claim 4 ,
wherein a washing fluid inlet is provided at the fluid inlet port or on the fluid inlet side other than the fluid inlet port for washing away unnecessary cells and other unnecessary substances remaining in the material for separating stem cell and
a cell recovering fluid inlet is provided at the fluid outlet port or at a site on the fluid outlet side other than the fluid outlet port for recovering cells captured by the material for separating stem cell.
9. The method of separating and recovering stem cells according to claim 8 ,
wherein a bag for receiving and storing a cell recovering fluid containing cells captured by the material for separating stem cell is provided at the fluid inlet port or washing fluid inlet or at a site on the fluid inlet side other than the fluid inlet port or washing fluid inlet.
10. The method of separating and recovering stem cells according to claim 9 ,
wherein the bag for receiving and storing a cell recovering fluid containing cells captured by the material for separating stem cell is a bag allowing cell culture therein.
11. A method of separating and recovering stem cells from a body fluid or a biological tissue-derived treated fluid according to claim 1 , wherein a mesh opening of the filter is 5 μm to 80 μm.