Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture.
Human adipose stromal cells (ASCs) reside within the stromal-vascular fraction (SVF) in fat tissue, can be readily isolated, and include stem-like cells that may be useful for therapy. An important consideration for clinical application and functional studies of stem/progenitor cells is their capacity to maintain chromosome stability in culture. In this study, cultured ASC populations and ASC clones were evaluated at intervals for maintenance of chromosome stability. Uncultured SVF (uSVF) cells were included for comparison. G-banded chromosome analysis demonstrated that ASCs are diploid and have a normal karyotype. However since only approximately 20 cells are examined, low levels of chromosome instability would not be detected. To increase detection sensitivity, fluorescence in situ hybridization was employed, to permit chromosome enumeration in larger numbers of interphase cells. Seven cultured ASC populations, two ASC clones and four uSVF samples were examined. Chromosome X and 17 probes identified diploid, tetraploid, and aneuploid interphase cells. Both cultured ASC populations [up to approximately 35 Population Doublings (PDs)] and uSVF cells exhibited a similar level of diploidy (97.8% n = 6,355 and 98.83% n = 1,197, respectively) and numerical abnormalities, suggesting that cultured ASCs are genomically stable and supporting their suitability for transplantation applications. In comparison, cultured primary human chorionic villus cells exhibited marked genomic instability resulting in an 11.6% tetraploidy rate after 8-10 PD. Thus effects of culture on genomic stability may be cell type dependent and should be tested by appropriately scaled interphase fluorescence in situ hybridization analysis in any ex vivo expanded cell population destined for transplantation.
Grimes
Brenda R
BR
Department of Medical and Molecular Genetics, Cytogenetics Division, Indiana University School of Medicine, 975 W. Walnut Street, Indianapolis, IN 46202, USA. brgrimes@iupui.edu
Steiner
Camie M
CM
Merfeld-Clauss
Stephanie
S
Traktuev
Dmitry O
DO
Smith
Danielle
D
Reese
Angela
A
Breman
Amy M
AM
Thurston
Virginia C
VC
Vance
Gail H
GH
Johnstone
Brian H
BH
Slee
Roger B
RB
March
Keith L
KL