A Novel Method to Isolate Mesenchymal Stem Cells from Bone Marrow in a Closed System Using a Device Made by Nonwoven Fabric

Abstract

Bone marrow stromal cells (BMSCs) include cells with multidirectional differentiation potential described as mesenchymal stem cells. For clinical use, it is important to develop a way to isolate BMSCs from bone marrow in a closed system without centrifugation. After screening 200 biomaterials, we developed a device containing a nonwoven fabric filter composed of rayon and polyethylene. The filter selectively traps BMSCs among mononuclear cells in bone marrow based on affinity, not cell size. The cells are then recovered by the retrograde flow. Using canine and human bone marrow cells, the biological properties of BMSCs isolated by the device were compared with those obtained by conventional methods using centrifugation. The total number isolated by the device was larger, as was the number of CD106(+)/STRO-1(+) double-positive cells. The cells showed osteogenic, chondrogenic, and adipogenic differentiation potential in vitro. Finally, the direct transplantation of cells isolated by the device without in vitro cultivation accelerated bone regeneration in a canine model of osteonecrosis in vivo. The proposed method is rapid and efficient, does not require a biological clean area, and will be useful for the clinical application of mesenchymal stem cells in bone marrow.

Mesenchymal stem cells and platelet lysate in fibrin or collagen scaffold promote non‐cemented hip prosthesis integration

“…[11][12][13] MSC and PRP alone or in combination have been used previously to promote bone regeneration. [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] Sheep have been demonstrated to be appropriate animals in orthopedic research due to their ability to tolerate surgery well and to recover quickly, as reported in our study and in other ones as well. 30,31 The majority of total hip replacement studies in sheep and goat pertain to the bone-cement interface, while the bone-prosthesis interface in non-cemented hip prostheses remains relatively unstudied.…”

“…MSC have been utilized in numerous studies as a source of osteogenic cells for bone repair because the bone‐specific differentiation of MSC has been well‐documented in vitro 11–13. MSC and PRP alone or in combination have been used previously to promote bone regeneration 14–29. Sheep have been demonstrated to be appropriate animals in orthopedic research due to their ability to tolerate surgery well and to recover quickly, as reported in our study and in other ones as well 30, 31.…”

Appropriate nonwoven filters effectively capture human peripheral blood cells and mesenchymal stem cells, which show enhanced production of growth factors

“…Thus, nonwoven filters in general may be suitable for the capture of cells for regenerative medicine. For example, nonwoven filters composed of rayon and polyethylene can be used to isolate bone marrow stem cells from bone marrow aspirates [4].…”

Composite nonwovens in medical applications

“…Isolation of these stem cells from total marrow is usually conducted using a sucrose-based density gradient column, which is a time-consuming process. After screening 200 biomaterials, Ito et al (2010) selected rayon-polyethylene nonwoven fabric for the development of a fi lter for the collection of mesenchymal stem cells from bone marrow hematopoietic mononuclear cells. Mesenchymal stem cells adhered tightly to highly hydrophilic and rough surfaces as compared to hydrophobic and smooth surfaces, resulting in a rapid and effi cient method of cell isolation fi ltration.…”