High Cellular Concentration of Peripheral Blood Progenitor Cells During Cryopreservation Adversely Affects CFU-GM but Not Hematopoietic Recovery

Keung, Yi‐Kong; Cobos, Everardo; Morgan, David S.; Park, Mary; Dixon, Sharon; Wu, Ke; Park, Chan H.

doi:10.1089/scd.1.1996.5.73

Cited by 11 publications

(11 citation statements)

References 6 publications

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“…Furthermore, a preliminary report concluded that such pretreatment also reduces the resistance of the cells to cryopreservation injury (41). Recently, it was reported that the cell concentration of PB HPC in the freezing bag is correlated with CFU-GM recovery (39). In our study, however, the cell concentration was the same for both BM HPC and PB HPC.…”

Section: Discussioncontrasting

confidence: 68%

“…Although the difference in median recovery was not significant, others have demonstrated that lower recoveries of CFU-GM are found with PB HPC than with BM HPC (7,39). This difference in poststorage recovery CFU-GM recovery is unexplained.…”

Section: Discussionmentioning

confidence: 72%

“…This difference in poststorage recovery CFU-GM recovery is unexplained. It has been demonstrated that extensive prior chemotherapy reduces the number of CFU-GM in PB HPC (36,(39)(40)(41). Furthermore, a preliminary report concluded that such pretreatment also reduces the resistance of the cells to cryopreservation injury (41).…”

Section: Discussionmentioning

confidence: 99%

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Effects of Long-Term Storage at -90°C of Bone Marrow and PBPC on Cell Recovery, Viability, and Clonogenic Potential

Ayello¹,

Semidei-Pomales²,

Preti³

et al. 1998

Journal of Hematotherapy

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Autologous BM and PB HPC are usually stored from weeks to months until reinfusion after myeloablative chemotherapy. HPC have been stored for up to 16 months at -90 degrees C, using a mixture of 5% DMSO, 6% hydroxyethyl starch (HES), and 4% HSA as a cryoprotectant. Long-term storage (LTS) has usually entailed rate-controlled freezing using 10% DMSO and preservation in liquid nitrogen. The effects of LTS at -90 degrees C on the in vitro cell recovery, viability, and colony-forming unit-granulocyte macrophage (CFU-GM) clonogenic potential of autologous HPC that were not transplanted was studied. Sixteen BM and sixteen PB HPC had been cryopreserved for a median of 53 months (range 27-71) and 35 months (range 26-78), respectively. Samples of frozen HPC were thawed after 48 h, and the nucleated cell count, viability by trypan blue exclusion, and culture for CFU-GM were obtained. Following LTS, the cells were thawed and examined using the same assays. No difference in the median percentage recovery of nucleated cells was found in either the BM or PB HPC between the samples stored for 48 h and after LTS (5.73 x 10(9) versus 5.61 x 10(9) and 6.20 x 10(9) versus 5.78 x 10(9), respectively). In addition, no difference in median percentage viability was found in either the BM or PB HPC sampled at 48 h and at the end of LTS (75% versus 74% and 75% versus 76%, respectively). Finally, the median number of CFU-GM cultured from BM HPC at 48 h was 2.41 x 10(5) (range 0.33-11.01 x 10(5)) and at the end of LTS was 1.93 x 10(5) (range 0.32-10.55), representing a median recovery of 93% (range 19%-308%). Similarly, the median number of CFU-GM cultured from PB HPC was 1.66 x 10(5) (range 0-50.57) and at the end of LTS was 0.93 x 10(5) (range 0-44.9), representing a median recovery of 80% (range 36%-165%). This difference in percentage recovery was not significant (p = 0.514). There was poor correlation between the number of nucleated cells harvested and the percentage recovery of nucleated cells, cell viability, or CFU-GM for either the BM or PB HPC. Similarly, there was poor correlation between the number of CFU-GM in the harvest and their percentage recovery following LTS for both BM and PB HPC. Finally, there was poor correlation between the storage time of the BM or PB HPC and the percentage recovery of nucleated cells, cell viability, and CFU-GM. These data suggest that LTS of HPC at -90 degrees C is not associated with decreased recovery of nucleated cells or in vitro viability and is associated with only a modest decrease in clonogenic potential. This indicates that storage of HPC at -90 degrees C for periods in excess of 3 years is possible.

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Section: Discussioncontrasting

confidence: 68%

Section: Discussionmentioning

confidence: 72%

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Effects of Long-Term Storage at -90°C of Bone Marrow and PBPC on Cell Recovery, Viability, and Clonogenic Potential

Ayello¹,

Semidei-Pomales²,

Preti³

et al. 1998

Journal of Hematotherapy

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“…There have been some investigations concerning the effect of high cell density during cryopreservation. 9,24 Few data are available on the very low cell densities used in this study (range 0.72 ϫ 10 6 /ml to 28 ϫ 10 6 /ml). In previous murine studies, Dicke et al 25 found a significant deterioration of progenitor cell survival when marrow cells were frozen at densities of 0.5 ϫ 10 6 /ml.…”

Section: Discussionmentioning

confidence: 99%

Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment

Beaujean¹,

Bourhis

Bayle

et al. 1998

Bone Marrow Transplant

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Summary:Conventional hematopoietic stem cell cryopreservation methods use a DMSO concentration of 10%. However, cells manipulated ex vivo may require more refined freezing protocols adapted to the specific cell suspension. In this retrospective study, we evaluated the results obtained with CD34 ؉ cells purified from peripheral blood of 39 patients on the CEPRATE SC System and frozen in 7.5% DMSO with a view to transplantation. ؉ cells and CFU-GM were significantly related to hematological recovery. Our data suggest that purified CD34 ؉ cells can be successfully cryopreserved in 7.5% DMSO and may represent a first step in establishing freezing parameters for selected CD34 ؉ cells.

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“…Consequently, it is necessary that isolated hASCs be cryopreserved for future experiments; however, current techniques for expanding, preserving and inducing such cells leave room for improvement. While it has been reported that bone marrow-derived mesenchymal stem cells (BMSCs) can be cryopreserved without loss of viability or osteogenic potential (Kotobuki et al, 2004), it has been reported that cryopreservation can reduce the viability, proliferation and differentiation ability of CD34 + haematopoietic stem cells (de Boer et al, 2002;Keung et al, 1996;Stylianou et al, 2006). Furthermore, while cryopreservation of intact human adipose tissue has been achieved (Shoshani et al, 2001), studies suggest that optimal viability requires the presence of cryoprotectant agents and controlled rate freezing (Moscatello et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Culture effects of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on cryopreserved human adipose-derived stromal/stem cell proliferation and adipogenesis

Hebert

et al. 2009

J Tissue Eng Regen Med

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Previous studies have demonstrated that EGF and bFGF maintain the stem cell properties of proliferating human adipose-derived stromal/stem cells (hASCs) in vitro. While the expansion and cryogenic preservation of isolated hASCs are routine, these manipulations can impact their proliferative and differentiation potential. This study examined cryogenically preserved hASCs (n = 4 donors), with respect to these functions, after culture with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) at varying concentrations (0–10 ng/ml). Relative to the control, cells supplemented with EGF and bFGF significantly increased proliferation by up to three-fold over 7–8 days. Furthermore, cryopreserved hASCs expanded in the presence of EGF and bFGF displayed increased oil red O staining following adipogenic induction. This was accompanied by significantly increased levels of several adipogenesis-related mRNAs: aP2, C/EBPα, lipoprotein lipase (LPL), PPARγ and PPARγ co-activator-1 (PGC1). Adipocytes derived from EGF- and bFGF-cultured hASCs exhibited more robust functionality based on insulin-stimulated glucose uptake and atrial natriuretic peptide (ANP)-stimulated lipolysis. These findings indicate that bFGF and EGF can be used as culture supplements to optimize the proliferative capacity of cryopreserved human ASCs and their adipogenic differentiation potential.

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High Cellular Concentration of Peripheral Blood Progenitor Cells During Cryopreservation Adversely Affects CFU-GM but Not Hematopoietic Recovery

Cited by 11 publications

References 6 publications

Effects of Long-Term Storage at -90°C of Bone Marrow and PBPC on Cell Recovery, Viability, and Clonogenic Potential

Effects of Long-Term Storage at -90°C of Bone Marrow and PBPC on Cell Recovery, Viability, and Clonogenic Potential

Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment

Culture effects of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on cryopreserved human adipose-derived stromal/stem cell proliferation and adipogenesis

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