title: Freezing harvested hMSCs and recovery of hMSCs from frozen vials for subsequent expansion, analysis, and experimentation.
abstract: Human multipotential stromal cells (hMSCs) are easily isolated from bone marrow and can be expanded by up to 200-fold in culture. Cultures of hMSCs are heterogeneous mixtures of stem/progenitor cells and more mature cell types. The proportion of each cell type in a given culture depends on how the cells are maintained. To maintain their stem cell-like qualities, hMSCs should be plated at low seeding densities (60-150 cells/cm2), lifted when between 60% and 80% confluent and should not be expanded beyond 4-5 passages. Thus, it is useful to establish a frozen bank of early passage cells. hMSCs store well in vapor phase liquid nitrogen (LN2) and are easily recovered for further expansion. This chapter describes one method of establishing a bank of early passage hMSCs using a seed lot system and the subsequent recovery of hMSCs from frozen stocks. The recovered cells can then be harvested and used for analyses of identification, functionality, in vitro and/or in vivo experimentation, or further expanded.
authors: Center for Gene Therapy, Tulane University Health Sciences Center, New Orleans, LA, USA.
authors: Roxanne L
authors: RL
authors: Reger
authors: Margaret R
authors: MR
authors: Wolfe
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005615:Freezing; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology