Evaluation of the reactivity of apoptosis markers before and after cryopreservation in cord blood CD34(+) cells

Stem Cells and Development
Nicholas J GrecoGary Moroff

Abstract

Umbilical cord blood (CB) CD34(+) cells, on the basis of flow cytometry analysis, are comprised of multiple populations. In in vitro assays, only CD34(regular) FSC(high) cells are functional and low percentages of nonfunctional CD34(regular) FSC(low) cells were determined to be present in liquid-stored CB. Liquid-stored CD34(regular) FSC(high) cells prior to cryopreservation were judged to be functional by the formation of erythroid and myeloid colonies and transmigration assays. We have further evaluated the occurrence of apoptosis in CB CD34(+) cells using various apoptotic markers to understand better the influence of storage conditions that could be utilized with transplantation of CB. Of the CD34(regular) FSC(low) cells shown in the present study, 20-45% were labeled with the apoptotic reagents annexin-V, fluorescent caspase peptide substrates, and the anti-mitochondrial antibody APO2.7, but these cells were minimally stained with 7-aminoactinomycin-D (7-AAD). These apoptotic reagents identify different cellular targets, indicating the initiation of the apoptotic cascade prior to cryopreservation/thawing. Following cryopreservation and thawing, the apoptotic markers SYTO-16, tetramethyl rhodamine ethyl ester (TMRE), and 7-...Continue Reading

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Citations

Mar 1, 2009·Biopreservation and Biobanking·John M BaustJohn G Baust
Dec 21, 2007·Advanced Drug Delivery Reviews·Sadamitsu Asoh, Shigeo Ohta
Jul 14, 2011·Journal of Cellular Biochemistry·Kateřina KuželováBarbora Brodská
May 28, 2011·Journal of Neuroscience Research·Nikolaos MemosManousos Konstadoulakis
Jan 5, 2010·Organogenesis·John G BaustJohn M Baust
Mar 24, 2017·Biopreservation and Biobanking·John G BaustJohn M Baust
Nov 4, 2017·In Vitro Cellular & Developmental Biology. Animal·John M BaustJohn W Harbell
Aug 30, 2018·Stem Cells·Sehar AfreenMiriam Erlacher

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