Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture

Brenda R. Grimes, Camie M. Steiner, Stephanie Merfeld-Clauss, Dmitry O. Traktuev, Danielle Smith, Angela Reese, Amy M. Breman, Virginia C. Thurston, Gail H. Vance, Brian H. Johnstone, Roger B. Slee, Keith L. March

Research output: Contribution to journalArticle

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Abstract

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 ∼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 ∼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.

Original languageEnglish (US)
Pages (from-to)717-724
Number of pages8
JournalStem Cells and Development
Volume18
Issue number5
DOIs
StatePublished - Jun 1 2009

Fingerprint

Genomic Instability
Interphase
Stromal Cells
Chromosomal Instability
Diploidy
Population
Tetraploidy
Stem Cells
Fluorescence In Situ Hybridization
Blood Vessels
Clone Cells
Transplantation
Chromosomes, Human, 21-22 and Y
Chorionic Villi
Chromosomes, Human, Pair 17
Aneuploidy
Karyotype
Cell Count
Chromosomes
Fats

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

Cite this

Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture. / Grimes, Brenda R.; Steiner, Camie M.; Merfeld-Clauss, Stephanie; Traktuev, Dmitry O.; Smith, Danielle; Reese, Angela; Breman, Amy M.; Thurston, Virginia C.; Vance, Gail H.; Johnstone, Brian H.; Slee, Roger B.; March, Keith L.

In: Stem Cells and Development, Vol. 18, No. 5, 01.06.2009, p. 717-724.

Research output: Contribution to journalArticle

Grimes, BR, Steiner, CM, Merfeld-Clauss, S, Traktuev, DO, Smith, D, Reese, A, Breman, AM, Thurston, VC, Vance, GH, Johnstone, BH, Slee, RB & March, KL 2009, 'Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture', Stem Cells and Development, vol. 18, no. 5, pp. 717-724. https://doi.org/10.1089/scd.2008.0255
Grimes, Brenda R. ; Steiner, Camie M. ; Merfeld-Clauss, Stephanie ; Traktuev, Dmitry O. ; Smith, Danielle ; Reese, Angela ; Breman, Amy M. ; Thurston, Virginia C. ; Vance, Gail H. ; Johnstone, Brian H. ; Slee, Roger B. ; March, Keith L. / Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture. In: Stem Cells and Development. 2009 ; Vol. 18, No. 5. pp. 717-724.
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AU - Smith, Danielle

AU - Reese, Angela

AU - Breman, Amy M.

AU - Thurston, Virginia C.

AU - Vance, Gail H.

AU - Johnstone, Brian H.

AU - Slee, Roger B.

AU - March, Keith L.

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