Long‐term storage at −80°C of hematopoietic progenitor cells with 5‐percent dimethyl sulfoxide as the sole cryoprotectant

Galmés, Antonio; Besalduch, J.; Bargay, Judy; Novo, Andrés; Morey, Miguel; Guerra, José M.; Durán, Maria Antònia

doi:10.1046/j.1537-2995.1999.39199116897.x

Cited by 81 publications

(76 citation statements)

References 4 publications

(3 reference statements)

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“…Long-term observations in this study confirm previous data regarding the feasibility of cryopreserving stem cells in diminished DMSO concentrations; as low as 2.2% for stem cells cryopreserved in liquid nitrogen 15 or as low as 5% for stem cells frozen and stored at -80°C. 8,9,[16][17][18] This study also confirms that the elimination of HES as an extracellular cryoprotectant can be carried out within the cryopreservation time scale of less than 6 months without affecting long-term hematologic recovery. However, we recommend caution when considering longer periods of cryopreservation.…”

Section: Outcome and Long-term Follow-up Datasupporting

confidence: 65%

“…This protocol involved storage in the same mechanical freezer in solutions containing 5% or 10% DMSO as the sole cryoprotectant without HES. [7][8][9] Most studies seem to indicate that this simplified procedure is associated with a reduction in infusion toxicity and lower costs, with a similar hematopoietic reconstitution and clinical outcome to more standard protocols. However, there is little data regarding the long-term hematologic recovery and clinical course when such protocols are used.…”

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confidence: 99%

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Long-term hematologic reconstitution and clinical evaluation of autologous peripheral blood stem cell transplantation after cryopreservation of cells with 5% and 10% dimethylsulfoxide at 80 C in a mechanical freezer

Galmés¹,

Gutiérrez²,

Sampol³

et al. 2007

Haematologica

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We report the long-term evaluation over 12 years of a simplified technique for stemcell cryopreservation at -80ºC without rate-controlled freezing and with 5% (n=251) or 10% (n=47) DMSO as the sole cryoprotectant. Platelet recovery was greater in the 5% DMSO group while long-term hematologic recovery did not differ. Factors influencing a faster hematologic recovery were infusion of more than 2.7×10 6 /kg of CD34 + cells, 10% DMSO cryopreservation and G-CSF. We confirm that the procedure is feasible with a reduction in infusion-related toxicity from 60% using 5% DMSO. Differences in hematologic reconstitution were not clinically significant if a minimum of 1. 1 The toxic effects related to DMSO infusion are generally dose-related and while they are usually mild, they can become severe.2-4 HES is a relatively non-toxic drug but it is related with long-lasting pruritus 5 and osmotic nephrotoxicity. Cryopreservation protocols usually involve ratecontrolled freezing followed by the storage of the HSC in either the liquid or vapor phase of liquid nitrogen. These procedures are time consuming and require expensive computerassisted devices.

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Section: Outcome and Long-term Follow-up Datasupporting

confidence: 65%

mentioning

confidence: 99%

Long-term hematologic reconstitution and clinical evaluation of autologous peripheral blood stem cell transplantation after cryopreservation of cells with 5% and 10% dimethylsulfoxide at 80 C in a mechanical freezer

Galmés¹,

Gutiérrez²,

Sampol³

et al. 2007

Haematologica

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“…They did not directly compare the original and current results for each individual specimen, but the data indicated that the stem cells stored the longest had the lowest CD34 þ cell counts and CFU-GM. 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.…”

Section: Discussionmentioning

confidence: 99%

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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“…Recently, there have been several reports suggesting that cells can be successfully cryopreserved with minimal concentrations of DMSO [39][40][41][42][43][44][45][46][47][48][49][50][51]. For example, Zhao et al [46] showed that the use of 5% or 10% DMSO in combination with either 20% or 70% serum produced similar results during cryopreservation of fetal human liver CD34 + cells.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Thirumala

Gimble

Devireddy

2010

Stem Cells and Development

101

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Developing effective techniques for the cryopreservation of human adipose-derived adult stem cells (ASCs) could increase the usefulness of these cells in tissue engineering and regenerative medicine. To this end, we investigated the post-freeze/thaw viability and apoptotic behavior of Passage 1 (P1) adult stem cells (ASCs) in 11 different media: (i) the traditional media containing Dulbecco's modifi ed Eagle's medium (DMEM) with 80% fetal calf serum (FCS) and 10% dimethyl sulfoxide (DMSO), (ii) DMEM with 80% human serum (HS) and 10% DMSO, (iii) DMEM with 1% methyl cellulose (MC) and 10% of either HS or FCS or DMSO, and (iv) DMEM with 0%, 2%, 4%, 6%, 8%, or 10% DMSO. Approximately 1 mL (10 6 cells/mL) of P1 ASCs were frozen overnight in a −80°C freezer and stored in liquid nitrogen for 2 weeks before being rapidly thawed in a 37°C water bath (1-2 min of agitation), resuspended in culture media, and seeded in separate wells of a 6-well plate for a 24-h incubation period at 37°C. After 24 h, the thawed samples were analyzed by bright-fi eld microscopy and fl ow cytometry. The results suggest that the absence of DMSO (and the presence of MC) signifi cantly increases the fraction of apoptotic and/or necrotic ASCs. However, the percentage of viable cells obtained with 2% DMSO and DMEM was comparable with that obtained in freezing media with 10% DMSO and 80% serum (HS or FCS), that is, ~84% ± 5% and ~84% ± 8%, respectively. Adipogenic and osteogenic differentiation behavior of the frozen thawed cells was also assessed using histochemical staining. Our results suggest that post-thaw ASC viability, adipogenic and osteogenic differentiability can be maintained even when they are frozen in the absence of serum but with a minimal concentration of 2% DMSO in DMEM.

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Long‐term storage at −80°C of hematopoietic progenitor cells with 5‐percent dimethyl sulfoxide as the sole cryoprotectant

Abstract: HPCs can be cryopreserved at -80 degrees C with 5-percent DMSO and stored at -80 degrees C no longer than 6 months. A 5-percent DMSO concentration is comparable to a with 10-percent concentration in terms of recovery and MNC viability.

Cited by 81 publications

References 4 publications

Long-term hematologic reconstitution and clinical evaluation of autologous peripheral blood stem cell transplantation after cryopreservation of cells with 5% and 10% dimethylsulfoxide at 80 C in a mechanical freezer

Long-term hematologic reconstitution and clinical evaluation of autologous peripheral blood stem cell transplantation after cryopreservation of cells with 5% and 10% dimethylsulfoxide at 80 C in a mechanical freezer

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

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

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