Investigation of telomere length and umbilical cord blood hematopoietic stem cells colony-formingpotential in cryoconservation

Abstract

The study was focused on proliferative capacities ofhuman hematopoietic cord blood stem cells at two differentmethods of cryopreservation. The proliferative potential wasdetermined by measurement of telomere length and colonyformingcell assay. The telomere length was estimated bytechnique that combined flow cytometry and fluorescencein situ hybridization on Beckman Coulter Cytomics FS-500.We used the Telomere PNA Kit/FITC for Flow Cytometry formeasuring telomeric sequences as well as MethoCult® GFH4435 in colony-forming cell assays. The programmed PlanerCryo freezer and direct transfer into liquid nitrogen vapourwere used for cryopreservation of samples. The correlationbetween telomere length and capacity of cells to forming colonyafter thawing were established. The programmed freezingshowed advantage over freezing by direct transfer cellsinto liquid nitrogen vapour by better keeping of proliferativecapacity hematopoietic cells.

References

[1] Ballen K.K. New trends in umbilical cord blood transplantation. Blood 2005; 105(10): 3786-92.

[2] Абдулкадыров К.М. Заготовка, хранение и лабораторное те- стирование пуповинной крови. Гематология. Новейший справочник. М.: «Эксмо», 2004

[3] Абдулкадыров К.М., Романенко Н.А., Старков Н.Н. Получе- ние и клиническое применение периферических гемопоэтических стволовых клеток из пуповинной крови. Вопросы онкологии 2000; 46(5): 513-20.

[4] Куртова А.В., Зуева Е.Е., Фрегатова Л.М. Методы банкирова- ния пуповинной крови. Иммунология 2005: 577-91.

[5] Владимирская Е.Б., Майорова О.А., Румянцев С.А. и др. Био- логические основы и перспективы терапии стволовыми клетками. М: «Медпрактика-М», 2005.

[6] Ballen K. Cord blood banking review. Hematologist 2004.

[7] Румянцев А.Г., Масчан. А.А. Трансплантация гемопоэтиче- ских стволовых клеток у детей. Руководство для врачей. М.: МИА, 2003.

[8] Гришина В.В., Тимохина Е.В., Андреева Л.Ю. Система сбора и фракционирования стволовых клеток пуповинной крови. Вопросы гинекологии, акушерства и перинаталогии 2004; 3(6): 50-4.

[9] Ballen K., Barker J., Stewart S. et al. Collection and preservation of cord blood for personal use. Biol. Blood Marrow Transpl. 2008; 14: 356-63.

[10] Blackburn E.H. Telomeres and telomerase: their mechanisms of action and the effects of altering their functions. FEBS Lett. 2005; 579(4): 859-62.

[11] Pipes L., Tsang T., Peng S. et al. Harris telomere length changes after umbilical cord blood transplant. Transfusion 2006; 46: 1038-43.

[12] Lansdorp M. Telomeres, stem cells, and hematology. Blood 2008; 111: 1759-66.

[13] Notaro R., Cimmeno A., Tabarini D. In vivo telomere dynamics of human hematopoietic stem cells. PNAS USA 1997; 94: 13782-5.

[14] Honda S., Weigel A., Hjelmeland M. et al. Induction of telomere shortening and replicative senescence by cryopreservation. Biochem. Biophys. Research Comm. 2001; 282: 493-8

[15] Hunt J., Armitage E., Pegg E. Cryopreservation of umbilical cord blood: 1. Osmotically inactive volume, hydraulic conductivity and permeability of CD34. cells to dimethyl sulphoxide. Cryobiology 2003; 46: 61-75.

[16] Hunt J., Armitage E., Pegg E. Cryopreservation of umbilical cord blood: 2. Tolerance of CD34. cells to multimolar dimethyl sulphoxide and the effect of cooling rate on recovery after freezing and thawing. Cryobiology 2003; 46: 76-87.

[17] Mazur P. Freezing of living cells: Mechanisms and implications. Am. J. Physiol. 1984; 247: 125-42.

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