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PubMed1998

Effects on rapid cooling of small samples in quenching.

Rapid cooling of small samples is necessary both to cryofixation for electron microscopy and to vitrification for cryopreservation. Several effects on the cooling rates of small samples quenched into liquid nitrogen were studied, including the diameter of samples, the subcooling of liquid nitrogen, the quenching speed, and the quenching distance. The heat flux is 1.4 x 10(6) W/m2; the cooling rate is also up to 8200 K/s at the CHF point of boiling curves for sphere of diameter 0.287 mm quenching into subcooled liquid nitrogen. It is also found that if the time of sample moving inside the liquid nitrogen is not longer than the time required for forming stable vapor in the liquid, the quenching boiling heat transfer is not influenced by the quenching speed. Several equation for calculating heat flux of samples are also presented.

Cryopreservation↗

PubMed2026

Protective effects of seminal exosomes on cryopreserved sperm via inhibiting oxidative damage.

This study aimed to explore the protective effect of seminal plasma exosomes (SPEs) on human sperm structure and function during cryopreservation and its potential mechanism. The samples were divided into two groups: the control group was treated solely with sperm cryoprotectant before freezing, while the exosome group was supplemented with SPEs. After cryopreservation and thawing, sperm progressive motility, normal morphological rate, and survival rate were evaluated. Furthermore, PKH67 labeling experiments were performed, and oxidative stress markers as well as energy metabolism indicators in sperm were detected. Subsequent mechanism exploration was conducted via proteomic analysis and protein validation assays. This work reveals that adding SPEs at a concentration of 1 or 2 mg/ml effectively improves sperm progressive motility after cryopreservation. After supplementing with SPEs, sperm glucose levels are reduced and mitochondrial membrane potential is enhanced. Simultaneously, SPEs alleviate oxidative stress by decreasing reactive oxygen species (ROS) and DNA fragment index (DFI) while increasing superoxide dismutase (SOD) activity. Functional annotation of proteomics reveals that 14 of the differentially expressed proteins (DEPs) are associated with sperm motility. Enriched metabolic pathways related to sperm motility and sperm protein validation experiments indicate that the expression of MAPK, p-MAPK, and p-JNK proteins in sperm is higher in the Exosome group than in the Control group. This study provides important theoretical support for the application of SPEs in mitigating cryopreservation damage to sperm by enhancing antioxidant capacity. The specific mechanism may be mediated by the MAPK/p-JNK pathway.

Male↗

PubMed2002

Retrospective multicentre study on mechanical and enzymatic preparation of fresh and cryopreserved testicular biopsies.

BACKGROUND: Isolation of sperm suitable for ICSI from fresh or frozen-thawed testicular sperm extraction (TESE) can be facilitated by mechanical or enzymatic processing of the samples. METHODS: A retrospective multicentre study was initiated to compare these two approaches. Eleven German centres provided data on their TESE cycles performed during the period 1996/1997. Quality of retrieved sperm, fertilization rates of injected oocytes, embryo quality, resulting pregnancy rates and evolution of pregnancies were evaluated. RESULTS: The percentage of cycles with at least some motile sperm available for injection was higher after mechanical preparation. Independent of the preparation method, fertilization rates were higher for motile compared with immotile sperm or elongated spermatids in all groups and in general higher for cryopreserved versus fresh samples. Embryo quality was significantly better after injection of motile sperm for all treatments and in particular after enzymatic versus mechanical processing of biopsies. Pregnancy rates were identical for embryos derived from sperm prepared mechanically or enzymatically from fresh or cryopreserved testicular samples. The abortion rate (32/172, 18.6%) and the rate of multiple implantations (32/140, 22.9%) were not different from results reported in the literature for ICSI using ejaculated sperm. CONCLUSION: In this retrospective multicentre study, no unequivocal advantage of one over the other preparation method could be identified in 839 ICSI cycles using testicular sperm from 549 patients.

Abortion, Spontaneous↗

PubMed2004

Functional significance of the cell volume for detecting sperm membrane changes and predicting freezability in dog semen.

Due to the similarity of plasma membrane changes induced by capacitation and cryopreservation, the parameters describing sperm response to capacitating conditions can be used for evaluating the cryopreservation response in many animal systems. In dog sperm, the response of the total sperm population to ionophore treatment has been shown to be an indication of the freezability of semen samples. Another sperm functional characteristic decisive for cryopreservability is cell volume regulation, due to the generation of essential osmotic gradients across the plasma membrane during the freeze-thaw cycles. In the present study, cryopreservation-induced changes in the membrane functional integrity were examined by monitoring the osmotically induced response of cell volume and the response to an ionophore in live cell populations. Cell volume measurements were performed on Percoll-washed suspensions of freshly diluted and frozen-thawed dog spermatozoa. The proportion of live acrosome-reacted cells was evaluated by flow cytometry after incubation under capacitating conditions in the presence of the calcium ionophore, A23187. During freezing-thawing, significant membrane changes occurred related to the disturbance of volume control ability and the loss of a proportion of live acrosome-reacted cells (P < 0.05). There were significant differences between individuals with respect to the degree of functional and structural membrane changes after thawing. Significant correlations were found between acrosomal integrity and functional membrane integrity. When assessed in freshly diluted semen, these parameters correlated with those of frozen-thawed semen samples, pointing to the similarities between mechanisms of cryopreservation-related changes and those mechanisms that mediate changes in membrane permeabilities and in cell volume regulation. The detection of changes in the sperm plasma membrane by monitoring the sperm cell volume represents a simple, rapid and sensitive method to estimate sperm quality after the cryopreservation procedure. The individual variability in response to osmotic stress or to calcium ionophore treatment appears to reflect the subtle differences in the sperm membrane functionality which are crucial for the prediction of cryopreservability.

Animals↗

PubMed2000

In-vitro differentiation of germ cells from frozen testicular biopsy specimens.

In some men with germ cell maturation arrest, spermatogenesis can be resumed during in-vitro culture of testicular biopsy samples. In this study, we examined whether similar differentiation events can be induced in cultured germ cells from cryopreserved testicular biopsy specimens. Fresh and cryopreserved aliquots of the same testicular biopsy samples were cultured in medium supplemented with FSH and testosterone. After 24 and 48 h of culture, the progression of spermatogenesis and the percentage of Sertoli cells with DNA damage, detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL), were evaluated. Spermatogenesis progressed in a similar way in fresh and cryopreserved aliquots over the first 24 h of culture. However, in contrast to fresh aliquots, no additional progress of spermatogenesis was detected between the 24 and 48 h time points. The percentage of TUNEL-positive Sertoli cells in fresh aliquots showed only a moderate increase after 24 h of culture, whereas most Sertoli cells from cryopreserved aliquots became TUNEL-positive during the same culture period. These data show that limited progression of spermatogenesis can be achieved by culturing cryopreserved testicular biopsy specimens for 24 h, but no additional benefit can be expected from prolonging the culture beyond this time point.

Biopsy↗

PubMed1984

[Studies on the mycotic and bacterial risk of contamination and the use of nipagin in the artificial insemination of cryosperm].

Primary bacterial and mycological contamination was studied in random human ejaculates. After various phases of cryopreservation secondary microbiological contamination was investigated. Furthermore, semen samples of several donors were inoculated with suspension of different concentrations of yeasts and with the test bacteria Escherichia coli K12 and Staphylococcus aureus SG 511. Microbiological results were then compared before an after cryopreservation. The cryopreserving process was carried out with and without antibiotics to test the antibacterial effectiveness of antibiotics under these conditions. Moreover, we investigated the usability of a chemical preservative (Nipagin) at cryopreservation. The following results were obtained: 9.8 per cent of samples showed primary mycological contamination (1.9 per cent with Candida albicans). Cryopreservation reduced the concentration of fungi by more than 90 per cent on average. The bacteriological investigations have shown, that with one exception the semen was without pathogenetic bacteria. This situation was altered scarcely during the cryopreserving process (also by use of cryoprotective medium without antibiotics). Out of 25 ejaculates 11 had primary non-pathogenetic bacteria. Test bacteria inoculated into semen were not influenced by the addition of antibiotics to the cryoprotective medium (CPM). Nipagin prevented respectively reduced a bacterial or a mycological contamination of the CPM during a storage of 14 days dependent of nipagin concentration but reduced the motility of human spermatozoa after cryopreservation. The results suggest the conclusion to prefer a portionate storage without nipagin at a temperature of nearly + 1 degree C, thereby the microbiological contamination may be neglected.

Anti-Bacterial Agents↗

PubMed2026

Cryopreservation-induced proteomic alterations in Pêga donkey (Equus asinus) spermatozoa.

Semen cryopreservation is a key tool for assisted reproduction and genetic conservation, but its efficiency remains limited in donkeys, compromising post-thaw sperm quality. This study aimed to characterize the proteomic profile of Pêga donkey spermatozoa and to investigate molecular mechanisms associated with cryopreservation-induced impairment of sperm function. Semen samples were collected from Pêga jacks and evaluated for sperm motility and vigor before and after cryopreservation. Quantitative proteomic analysis was performed by LC-MS/MS, followed by bioinformatic characterization of differentially abundant proteins. Cryopreservation markedly reduced sperm motility in all animals, whereas sperm vigor showed only a non-significant tendency toward reduction, suggesting sublethal cryoinjury primarily affecting flagellar efficiency. Proteomic profiling identified 554 proteins, of which 98 were differentially abundant between in natura and cryopreserved spermatozoa. Functional enrichment analyses showed that these proteins were mainly associated with energy metabolism, mitochondrial oxidative phosphorylation, glycolysis, cytoskeletal organization, signal transduction, proteostasis, and oxidative stress response. Notably, proteins involved in ATP production, mitochondrial function, and axonemal organization were significantly altered, supporting a mechanistic link between metabolic dysfunction, flagellar structural disorganization, and reduced post-thaw motility. Overall, cryopreservation induced coordinated and compartment-specific remodeling of the donkey sperm proteome, particularly affecting pathways essential for motility and functional competence. These findings provide new molecular insights into the cryobiological vulnerability of donkey spermatozoa and establish a mechanistic basis for the development of more effective, biology-driven cryopreservation strategies for this species.

Animals↗

PubMed1983

Production of genetically identical sets of mammals: cloning?

In the past few years, there has been phenomenal progress in producing genetically identical mammalian multiplets by asexual means. The simplest method is to divide embryos into two, three, or four parts, and transfer the parts to surrogate mothers for gestation to term. Another approach is to inject nuclei from totipotent embryonic cells into fertilized one-cell embryos and to remove both male and female pronuclei. When these embryos reach the blastocyst stage, nuclei from their totipotent cells can be removed and injected into one-cell embryos, thus amplifying the process ad infinitum. Subsets of these embryos can be stored in liquid nitrogen to make genetic material for additional copies available indefinitely. Some of these techniques are fairly efficacious, others are not. No reliable techniques are available for making genetic copies of nonhomozygous adult mammals (or other vertebrates) unless samples of embryonic cells were cryopreserved when the individuals were embryos. However, injection of nuclei of spermatogonia into enucleated one-cell embryos is a promising strategy. The genetic totipotency of these nuclei is obvious since they become gamete nuclei. Moreover, the birth of mice from the diploidized genetic material of a single X-bearing spermatozoan after removal of the female pronucleus from a one-cell fertilized embryo suggests that we should eventually be able to make genetic copies of adult male mammals. This approach would not work for adult females because their oogonia have all become oocytes, and the DNA configuration in oocytes is not identical to somatic cells due to meiotic crossing over.

Animals↗

PubMed1995

Human allograft vein failure: immunohistochemical arguments supporting the involvement of an immune-mediated mechanism.

The aim of this study was to search for signs suggestive of an ongoing immune-mediated reaction in failed human cryopreserved venous allografts. In 15 samples, the authors analyzed: (1) the pattern of morphological changes; (2) the density, distribution, and phenotype of leukocytic infiltrate; and (3) the expression of class II major histocompatibility complex (MHC) antigens and inducible adhesion molecules. Two groups of samples could be recognized. In samples explanted before 3 months after grafting, the structure of the vessel wall was preserved. A dense leukocytic infiltrate was present within the intima and around the numerous vasa vasorum located in medial and adventitial layers. Class II MHC antigens and cytokine-dependent molecules were induced on endothelial cells lining the vasa vasorum and on residual smooth muscle cells. In samples explanted after 3 months of evolution, the vessel wall has lost its normal structure and contained few vasa vasorum. A few leukocytes were detected around capillary vessels located in the peripheral connective tissue surrounding the graft. Class II MHC antigens and adhesion molecules were induced on endothelial cells lining the peripheral capillary vessels. These results suggest the involvement of an immune-mediated mechanism at the early stage of the evolution of failed human venous allografts.

Cell Adhesion Molecules↗

PubMed2003

Association between antenatal cytokine production and the development of atopy and asthma at age 6 years.

BACKGROUND: Various lines of evidence suggest that antenatal factors are important in determining susceptibility to atopy and asthma. One possible mechanism is cytokines, production of which in the placenta is high throughout gestation and which protect placental integrity via control of local immunological homoeostasis. We investigated antenatal cytokine concentrations in a prospective birth cohort, intensively monitored for atopy and asthma outcomes at age 6 years. METHODS: Cryopreserved cord-blood serum samples from 407 children were assayed for interleukins 4, 5, 6, 10, 12, and 13, interferon gamma, and tumour necrosis factor alpha (TNFalpha). Associations between family, antenatal, and perinatal factors, cord-blood cytokine concentrations, and atopy or asthma outcomes were analysed by logistic regression. Causal effects of cytokines on outcomes were estimated by propensity scores based on family, antenatal, and perinatal factors. FINDINGS: Detectable cord-blood concentrations of interleukin 4 and interferon gamma were each associated with lower risk of physician-diagnosed asthma (adjusted odds ratios 0.60 [95% CI 0.37-0.99] and 0.60 [0.37-0.97] respectively), current asthma (0.59 [0.33-1.00] and 0.39 [0.22-0.71]), and current wheeze (0.55 [0.32-0.93] and 0.52 [0.31-0.90]) and atopy (sensitisation to some inhalant allergens) outcomes at 6 years. High concentrations of TNFalpha were associated with lower risk of atopy but not with asthma risk. These associations were broadly unaltered by propensity-score adjustment. Maternal smoking was associated with higher risk of both wheeze at 6 years and lower concentrations of interleukin 4 and interferon gamma in cord blood. INTERPRETATION: The mechanism underlying attenuated T-helper-1/T-helper-2 cytokine production in high-risk children also apparently operates in control of cytokine production in the fetoplacental unit. The finding that this mechanism is dysregulated by maternal smoking suggests it is a target for antenatal environmental factors relevant to asthma aetiology.

Asthma↗

PubMed2003

Anti-inflammatory and varicocele treatment in nonobstructive azoospermia.

A 34-year-old male patient with nonobstructive azoospermia and childlessness for more than 2 years is presented. As even severely damaged testes often harbour cryptic areas with remaining spermatogenesis, such cases might be suitable for spermatid extraction from bilateral testicular biopsies. Taking into account that testicular biopsies bear the risk of permanent testosterone insufficiency, less invasive andrological treatment modalities were considered. Under the assumption of frequent silent testicular inflammatory reactions, the nonsteroidal anti-inflammatory drug ibuprofen was administered. Additionally, a concomitant left-sided varicocele was removed by retrograde sclerotherapy. Within 1 year, the spermiogram showed spermatozoa at a concentration of 0.6-1.2 million ml(-1). This allowed ejaculate samples to be stored for cryopreservation to enable in vitro fertilization with intracytoplasmic injection of spermatozoa. Our successfully performed anti-inflammatory and varicocele treatment underlines the value of conservative andrological therapies. Despite the modern possibilities of assisted reproductive procedures the clinical andrologist should be aware of a careful andrological examination and of less invasive therapeutic options such as anti-inflammatory treatment and varicocele repair.

Adult↗

PubMed1993

Potential genetic risks of using semen collected during chemotherapy.

The results of many studies on the effects of mutagens on germ cells of the testes indicate that the recommendation of Carson et al. (Hum. Reprod., 6, 992-994, 1989) that semen samples collected during chemotherapy and cryopreserved may be safely used for insemination is incorrect because of the large potential for genetic risk to the offspring. Data from experimental animals, which indicate high levels of mutagenic effects in offspring from matings during or soon after treatment of the male with chemotherapy or radiation, are reviewed. The relevance of these data to the human situation is discussed. Based on these results, I recommend that patients not only complete their semen cryopreservation prior to the initiation of chemotherapy, but that they practice reliable contraception from the time of initiation of treatment until 6 months after completion of treatment.

Animals↗

PubMedDate not supplied

Fluorescent-activated cell-sorting analysis of intracellular interferon-gamma and interleukin-4 in fresh and frozen human peripheral blood T-helper cells.

T-helper (Th) cells can be classified into at least three subsets based on their cytokine profiles: Th0, Th1, and Th2. The functional significance of each subset of Th cells can be determined in isolated human peripheral blood mononuclear cells (PBMC). Using two- or three-color cytometric detection of intracellular cytokines. These analyses have been limited by the requirement for fresh cells making sequential samples and longitudinal studies difficult. Cryopreservation of PBMC in liquid nitrogen for up to 1 year was evaluated to determine whether the Th1/Th2 ratio remained unchanged in cryopreserved lymphocytes. Aliquots of human PBMC from normal volunteers analyzed for activation using phorbol myristate acetate and evaluated using morphology showed that the surface marker expression was unchanged in fresh and frozen cells. Cytokine expression was measured using intracellular cytokine staining and three-color flow cytometric analysis. The percentages of cells producing interferon (IFN)-gamma or interleukin (IL)-4 were determined after 16 hours of phorbol myristate acetate and ionomycin stimulation in the presence of brefeldin A. No significant difference was found in cytokine production between fresh and frozen cells. The percentage of IFN-gamma and IL-4 producing CD3-positive fresh T cells was 19.2+/-5.8 percent and 0.9+/-0.4 percent vs. 17.6+/-0.75 percent and 0.9+/-0.3 percent, respectively, for frozen PBMC. The effects of thermal injury on the Th1/Th2 cytokine ratio and the development of hypertrophic scarring were then determined. Twelve burn patients examined 4 weeks postburn showed a significant shift in the Th1/Th2 ratio, compared with 13 normal human volunteers used as controls. IL-4 levels in the patient group were significantly higher than controls at 1 month postburn (12.7+/-2.6 percent vs. 3.9+/-0.5 percent, p<0.01) and IFN-gamma levels were significantly lower (9.3+/-1.7 percent vs. 15.3+/-2.3 percent, p<0.05). Thus, PBMC can be cryopreserved for up to 1 year, enabling investigation of chronologic changes in Th1/Th2 profiles. It is suggested that a "locked on" Th2 profile may contribute to the development of hypertrophic scarring after burn injury.

Adult↗

PubMed1991

Supernatant cell counts from long-term bone marrow culture correlate with the speed of engraftment following autologous bone marrow transplantation.

This study evaluates the relationship between bone marrow growth in a long-term bone marrow culture (LTBMC) system and speed of engraftment of the same marrow following autologous bone marrow transplantation (ABMT). Bone marrow from 21 patients transplanted with unmanipulated, non-cryopreserved autologous marrow was cultured. Samples from 21 normal donors were cultured to establish the normal supernatant cell count range. Supernatant counts from LTBMCs established from marrow taken from patients at the time of bone marrow harvest were compared with the time to neutrophil and platelet engraftment. Supernatant counts, particularly after 1 week in culture, showed close correlation with time to neutrophil and platelet engraftment following ABMT (r = 0.733, p less than 0.01; r = 0.735, p less than 0.01 respectively). Where supernatant cell counts were within the normal range rapid engraftment was predicted (neutrophils greater than 0.5 x 10(9)/l within 21 days, platelets greater than 50 x 10(9)/l within 28 days) and if supernatant counts were below this range, engraftment was predicted to be delayed. After 1 week in culture, the speed of neutrophil and platelet engraftment were correctly predicted in 19 and 18 cases respectively. Preliminary data suggest that LTBMC of marrow obtained 2-6 weeks before harvesting provides similar data, thus allowing the opportunity to intervene, for example with growth factors, in selected patients.

Adolescent↗

PubMed1987

[Use of human albumin as an additional cryoprotective agent in freeze preservation of hematopoietic stem cells].

14 bone-marrow samples of healthy donors were cryopreserved with 5% of DMSO in combination with different volume percentages of 20% human albumin, with the protective impact of the respective freezing mixture on the proliferating capacity of early hemotopoietic precursor cells being determined by means of CFU-c-technique. A combination of 5% of DMSO with 20% of human albumin with an addition of 20% of autologous serum proved to be the most favourable freezing protection. CFU-c-recovery which was achieved in this way amounted to 78 +/- 7%. A protective impact of human albumin for CFU-c during incubation with the freezing medium at room temperature was statistically not significant. Thus, the method presented is suitable for cryopreservation of bone marrow for autologous transplantation. The advantages over the traditional procedure results from the slight degree of toxicity by reducing the DMSO percentage from the common 10% to 5% as well as from the fact that the thawed cell suspension may be directly infused with the substance for freezing protection, thus avoiding an additional loss of proliferative material. The procedure may be used especially for autologous transplantation in childhood.

Adult↗

PubMed1988

Cryopreservation of isolated blood vessels.

Canine saphenous veins were immersed in fetal calf serum (FCS) containing various cryoprotective agents, slowly frozen and stored for several weeks at subzero temperatures. Pharmacological investigations of frozen/thawed tissues revealed considerable attenuation of the contractile force of frozen stored veins as compared to unfrozen veins. The best recovery after thawing of frozen stored canine veins was obtained on tissues which had been frozen slowly to -70 degrees C and stored in liquid nitrogen while being immersed in FCS containing 1.8 mol/l dimethyl sulfoxide (DMSO). Though the maximum response to noradranline of helical strips prepared from these veins was diminished to about 60% the evidence suggests that there may be a very good preservation of the main biochemical properties, such as monoamine oxidase activity, endogenous prostaglandin synthesis and neuronal uptake mechanism in veins stored under these conditions. The same method of cryopreservation was applied to store samples of human veins. Comparison of the pD2 values for various agonists and of the blocking activities of various antagonists of both 5-HT receptors and alpha-adrenoceptors yielded an excellent correlation between the parameters determined on frozen/thawed and unfrozen human veins. It is concluded that freezing isolated blood vessels may be considered an effective means of preserving and storing vascular tissues for pharmacological investigations.

Animals↗

PubMed1996

The variation of Pf155/RESA gene in field isolates of Plasmodium falciparum from Thailand.

Pf155/RESA, an antigen found on the surface of Plasmodium falciparum red blood cell membrane was once a proposed malarial vaccine candidate. The complete sequence of Pf155/RESA gene from one strain and partial sequence from two other isolates revealed that the gene is well conserved. But polymorphism of other antigenic encoded regions occurs with high frequency among isolates especially those collected from the field. Using solid phase sequencing technique, the nested PCR products of upstream 3' repeated region of exon 2 RESA gene were studied in 150 P. falciparum isolates. Of which 117 isolates were directly collected from the field and sequenced. Other samples studied include clones and cryopreserved of previously cultured isolates. The resulting sequences are compared with previously existing data of F32 (Tanzania) and FC27 (Papua New Guinea) designated as allelic type I and II respectively. Sequence analysis of the 150 P. falciparum showed that the amplified region of RESA gene was highly variable with substitution ranging from one to six bases and these allelic variables can be divided into 10 types. The frequency of type I(F32) occurrence is 70.86 percent, type III 13.38 per cent and 0.78 percent to 5.51 per cent for others. As a result of allelic polymorphism, the amino acid sequence is highly variable and this may cause Pf155/RESA to be an inefficient antigen.

Alleles↗

PubMed1998

Evaluation of the gemini infusion pump for the safe delivery of peripheral blood progenitor cells (stem cells).

The purpose of this in vitro study was to determine whether the Gemini PC-2TX infusion pump could safely deliver peripheral stem cells (PSC) for an autologous PSC transplant. For purposes of hypothesis testing, it was assumed that there would be no significant difference in CD34+ cell counts and colony-forming units-granulocyte, macrophage (CFU-GM) when the PSCs were administered by an IMED PC-2TX infusion pump as opposed to an intravenous push method. The American Red Cross collected 50-ml samples of PSCs from four donors by apheresis. These cells were tested for CD34+ using flow cytometry and for functional progenitor cells using a CFU-GM assay. The cells were cryopreserved after testing. For our study, samples were tested simultaneously at a single facility. Each sample was individually thawed and a baseline thaw sample collected; 10 ml of the donor specimen was pushed through a syringe into a specimen container (intravenous push sample). The remainder of the specimen was infused through the IMED Gemini PC-2TX pump into a specimen container (intravenous pump sample). All samples were assayed for CD34+ cell counts and CFU-GM. Data analyses were conducted using the t-test for paired samples, with values of P < 0.05 considered significant. Results failed to demonstrate a statistically significant difference between the CD34+ or CFU-GM results of the intravenous push and intravenous pump specimens. Additionally, we failed to find a statistically significant difference when we compared the intravenous push and the intravenous pump specimens with the baseline thaw sample. The results of this study support the hypothesis that the Gemini PC-2TX infusion pump can safely deliver PSCs for the purposes of stem cell transplantation.

Antigens, CD34↗