Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing

Cilloni, Daniela; Garau, D.; Regazzi, Ester; Sammarelli, Gabriella; Savoldo, Barbara; Caramatti, Cecilia; Mangoni, L; Rizzoli, Vittorio; Carlo‐Stella, Carmelo

doi:10.1038/sj.bmt.1701601

Cited by 34 publications

(39 citation statements)

References 26 publications

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“…Other studies have also shown a decrease in CFU-GM or engraftment with long-term storage but are less applicable because of the use of less extreme freezing temperatures 17 or the use of animal models 18 and studies that report no decrease in SCP markers have been limited by relatively short median storage times. [9][10][11][12]19 In this study, recovery of SCP, TNC, cell viability and total CD34 þ cell count decreased as storage time, up to 19 years, increased. No previous study has analysed stem cells stored for this long and no previous study, using PB or BM stem cells, has directly compared individual samples results from the time of original harvest with the current time post long-term storage.…”

Section: Discussionmentioning

confidence: 97%

Relative recovery of haematopoietic stem cell products after cryogenic storage of up to 19 years

Fernyhough

Buchan

McArthur

et al. 2012

Bone Marrow Transplant

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There is an increasing trend towards long-term frozen storage of haematopoietic stem cells. For such stem cells, harvested from peripheral blood (PB) or BM, it is not known if stem cell viability decreases with time. In this study, 31 separate bags of stem cell product (SCP) stored for 11-19 years (median 15 years) were assessed for total nucleated cell (TNC) count, colony forming unitgranulocyte/macrophage (CFU-GM), CD34 þ cell count and cell viability. The results were compared with the initial results obtained for the products at the time of stem cell harvest, and the percentage recovery of each parameter was plotted against time. Recovery of TNC, CD34 þ cell count and cell viability decreased with time (P ¼ o0.01) but CFU-GM did not. This study shows that SCPs harvested from PB and BM do deteriorate with long-term storage. This could have an impact on rates of engraftment.

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Section: Discussionmentioning

confidence: 97%

Relative recovery of haematopoietic stem cell products after cryogenic storage of up to 19 years

Fernyhough

Buchan

McArthur

et al. 2012

Bone Marrow Transplant

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“…Specifically, fast cooling forms intracellular ice crystals, which results in cell destruction and slow cooling forms ice crystals in the extracellular space, with consequent cellular dehydration. Selection of a cryoprotectant, as well as a suitable freezing rate serves to protect cells from these adverse effects (3,4).…”

Section: Introductionmentioning

confidence: 99%

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Martinetti¹,

Colarossi²,

Buccheri³

et al. 2017

Biomedical Reports

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Abstract. Stem cells are an important tool for the study of hematopoiesis. Despite developments in cryopreservation, post-thaw cell death remains a considerable problem. Cryopreservation protocol should limit cell damage due to freezing and ensure the recovery of the functional cell characteristics after thawing. Thus, the use of cryoprotectants is essential. In particular, the efficacy of trehalose has been reported for clinical purposes in blood stem cells. The aim of the current study was to establish an efficient method for biological research based on the use of trehalose, to cryopreserve pure peripheral blood stem cells. The efficacy of trehalose was assessed in vitro and the cell viability was evaluated. The data indicate that trehalose improves cell survival after thawing compared with the standard freezing procedure. These findings could suggest the potential for future trehalose application for research purposes in cell cryopreservation. IntroductionStem cells in biological research provide a significant source of information and, thus, contribute to the development of novel therapeutic strategies. In particular, the ability to cultivate stem cells and human hematopoietic progenitor cells in vitro is the fundamental basis for investigating hematopoiesis. An improved understanding of the mechanisms that regulate cell proliferation and differentiation during the stages of hematopoiesis would further elucidate the molecular characteristics of diseases (which are characterized by excessive expansion or a functional defect of certain immature blood components), and facilitate the identification of substances that are able to specifically protect healthy cells from the action of cytotoxic drugs.Hematopoietic stem and progenitor cells (HSPCs) may be isolated from peripheral blood (PB), bone marrow (BM) or umbilical cord blood (CB) (1).Although significant improvements to cell cryopreservation procedures have been achieved (2), the improvement of current cryopreservation protocols that lead to a high mortality rate after thawing for the crystal formations that arise during freezing is a primary goal. Specifically, fast cooling forms intracellular ice crystals, which results in cell destruction and slow cooling forms ice crystals in the extracellular space, with consequent cellular dehydration. Selection of a cryoprotectant, as well as a suitable freezing rate serves to protect cells from these adverse effects (3,4).Cryoprotectants are divided into two classes: Penetrating and nonpenetrating (5). The penetrating cryoprotectants include glycerol and 1,2-propanediol and dimethyl sulfoxide (Me 2 SO); the latter is commonly used for HSPC cryopreservation (6).The non penetrating cryoprotectants comprise polyvinyl pyrrolidone, trehalose, fructose, sucrose and glucose.Trehalose is a non-toxic disaccharide of glucose that preserves the structural integrity of the cells during freezing and thawing (7). Specifically, trehalose is found in numerous organisms, such as nematodes and yeasts, which are capable of surviving du...

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“…Unfortunately, this technique requires sophisticated, costly equipment and a certain personal expertise, which are not available at every center (Almici et al, 2003;Perez-Oteyza et al, 1998). Several single armed studies have reported excellent viability and hematopoietic recovery rates with uncontrolled freezing procedures in bone marrow (Clark, Pati, & D. McCarthy, 1991;Stiff, Murgo, Zaroulis, DeRisi, & Clarkson, 1983) and peripheral blood progenitor specimens (Almici et al, 2003;Cilloni et al, 1999;Halle et al, 2001). Few studies directly compared outcomes of controlled rate to uncontrolled rate freezing processes.…”

Section: Alternatives To Standard Dmsomentioning

confidence: 99%

“…Temperatures range from -196°C to -80°Celsius (Aird, Labopin, Gorin, & Antin, 1992;Cilloni et al, 1999;Galmés et al, 1999;Halle et al, 2001;Rubinstein et al, 1995;Son, Heo, Park, H. H. Kim, & K. S. Lee, 2010;Valeri & Pivacek, 1996). The initially used storage temperatures of -196°C, reflecting the liquid phase nitrogen storage, have largely been replaced by temperatures of -156°C to -135°C, reflecting the vapor phase storage (Berz, McCormack, Winer, Colvin, & Peter J Quesenberry, 2007) in BM-, PB-as well as umbilical cord derived stem cells(Aird, Labopin, Gorin, & Antin, 1992;Cilloni et al, 1999;Galmés et al, 1999;Halle et al, 2001;Rubinstein et al, 1995; Son, Heo, Park, H. H. Kim, & K. S. Valeri & Pivacek, 1996). This shift in culture was mainly induced by the observation that infectious pathogens can survive and be propagated in the liquid nitrogen phase (Bielanski & Vajta, 2009).…”

Section: Storage Temperaturementioning

confidence: 99%

Cryopreservation of Hematopoietic and Non-Hematopoietic Stem Cells – A Review for the Clinician

Berz¹,

Colvin²

2012

New Advances in Stem Cell Transplantation

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Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing

Cited by 34 publications

References 26 publications

Relative recovery of haematopoietic stem cell products after cryogenic storage of up to 19 years

Relative recovery of haematopoietic stem cell products after cryogenic storage of up to 19 years

Effect of trehalose on cryopreservation of pure peripheral blood stem cells

Cryopreservation of Hematopoietic and Non-Hematopoietic Stem Cells – A Review for the Clinician

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