Magnetic stromal layers for enhanced and unbiased recovery of co-cultured hematopoietic cells.
Cell co-culture systems have a long history of application in hematology and hold promise for successful hematopoietic stem and progenitor cell expansion. Here we report that various types of stromal cells used in such co-cultures can be rapidly and efficiently labeled with l-lysine-modified Fe3O4 magnetic nanoparticles. Hematopoiesis-supporting activity does not seem to be compromised after magnetic labeling of stromal cells, and the loss of the label by stromal layers during extended culturing is negligible. Magnetic labeling allows for simple and efficient removal of stromal component, yielding unbiased hematopoietic cell populations. When Lin(-) bone mouse marrow fraction was co-cultured with magnetic stromal layers and resulting cell populations were harvested by trypsinization, the yields of total nucleated cells, colony forming cells, and phenotypically primitive Lin(-)Sca-1(+)c-kit(+) subset were substantially higher as compared with nonadherent cell fractions harvested after conventional stromal co-culture. The advantage offered by the magnetic stroma approach over the traditional one was even more significant after a second round of co-culture and was more dramatic for more primitive hematopoietic cells. We conclude that magnetic stromal layers represent a simple, efficient, and convenient tool for co-culturing and subsequent recovery of sufficiently pure unbiased populations of hematopoietic cells.
Engelhard Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia.
Maria V
MV
Savvateeva
Postovsky Institute of Organic Synthesis, Russian Academy of Sciences (Ural Branch), Ekaterinburg, 620990, Russia.
Alexander M
AM
Demin
Postovsky Institute of Organic Synthesis, Russian Academy of Sciences (Ural Branch), Ekaterinburg, 620990, Russia.
Victor P
VP
Krasnov
Engelhard Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia. Electronic address: abelyavs@yahoo.com.
Alexander V
AV
Belyavsky
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout
D058185:Magnetite Nanoparticles
Adhesion; Bone marrow; Co-culture; Magnetic isolation; Nanoparticle; Stem cells