An alternative test for determining autologous stem cell transplantation cell harvesting outcome: Comparison between predictive values of two tests.

Daily CD34+ cells enumeration as a success indicator of stem cell pheresis procedure using flow cytometry is costly, lengthy, and labor-intensive. Thus, finding a simpler method to achieve the optimum time for harvesting the minimum required stem cells for transplantation could be helpful. The aim of this study was to evaluate the predictive value of reticulocytes fractions and their sensesivity and specificity in guiding CD34+ cell harvesting by G-CSF mobilization strategy. In this study, 49 candidates for autologous peripheral blood stem cell transplantation were enrolled. Before leukapheresis, the immature reticulocytes fraction (IRF) and CD34+ cell count were measured. Moreover, patients were evaluated for leukapheresis outcomes in two MNC and cMNC groups. Here we demonstrated that IRF, LFR, and MFR with the associated criterion of >17.3, ≤82.5, and >15.9, respectively, earned 100 % specificity and 47.2 %, 47.22 %, and 41.46 % sensitivity to predict the minimum required CD34+ cell count. Furthermore, IRF-V (Value) and MFR-V with the associated criterion of >0.77 and >0.55, respectively, earned 58.33 %, 66.67 % sensitivity and 84.62 %, 69.23 % of specificity, separately. As only MFR-V was able to predict the platelet engraftment (P-value = 0.014), none of the other above mentioned factors were not able to predict the neutrophil engraftment. Likewise, it was shown that patients who underwent MNC leukapheresis had a statistically significantly higher total WBC, harvested CD34+ cells, MNCs/ kg, and lower apheresis durations (P-values<0.05). Taken together, using IRF and its maturity stages seems to be a compelling predictor of minimal required CD34+ cells in autologous peripheral blood stem cell transplantation.

Effects of mobilized peripheral blood stem cells on treatment of primary lower extremity lymphedema.

OBJECTIVE: Lymphedema is a chronic debilitating disease characterized by the accumulation of fluid in the extremities as a result of lymphatic system impairment. Current treatments fail to restore the functionality and structural integrity of the lymphatic vessels lost in this condition. In this study, autologous mobilized peripheral blood stem cell transplantation was used and its potential efficacy and safety were evaluated in treating this condition. METHODS: Ten patients with primary lymphedema in the lower extremity received granulocyte-colony stimulating factor subcutaneously for 4 days, to stimulate stem cell mobilization, after which 200 to 250 mL of blood was drawn from each patient and used to collect stem cells. Mobilized stem cells were counted by flow cytometry with International Society of Hematotherapy and Graft Engineering method. In two sessions, 3 weeks apart, these stem cells were injected subcutaneously in the affected limb at approximately 80 points, along the lymphatic vessels. Each patient was followed for 6 months, during which changes in the limb volume and circumference were measured. Lymphangiogenesis was evaluated by biopsy, the lymphoscintigraphic transport index was calculated using Lymphoscintigraphy, and quality of life was surveyed. RESULTS: In this study, patients received on average 9.5 ± 6.8 × 108 mononuclear cells (which divided into 2 × 106 CD34+ cells for each session) in two sessions. The volume of the lower limbs decreased in 60% of patients. One patient showed a slight increase in the volume of lower limbs and three showed no change. The average limb volume was 4469.41 ± 1760.71 cm3, which on average differed from the average initial limb volume by 232.88 ± 392.53 cm3. Quality of life was reported as slightly increased in 60% of patients. The lymphoscintigraphic transport index suggested improvement in 60% of the patients. Likewise, tissue samples showed a 60% increase in lymphatic vessels. CONCLUSIONS: Subcutaneous injection of autologous hematopoietic stem cells harvested from peripheral blood into patients with primary lower limb lymphedema is feasible, potentially effective, and without serious adverse effects. However, a larger scale study with more patients is needed to validate our results. Last, to increase the effectiveness of this treatment, the optimal dose of cells injected and the requirement for additional growth factors need further study.

Peripheral blood stem cell apheresis in low-weight children: A single centre study.

Peripheral blood stem cell transplantation (PBSCT) is now widely used in both malignant and non-malignant hematologic diseases as a treatment strategy. Using this approach, a controversial group of donors is children weighing 20 kg or less. The aim of this study was to evaluate results of allogeneic and autologous PBSCT and also the efficacy of our suggested alternative method for a custom prime in cell harvesting of this group. All the participants' demographic and laboratory data were collected before apheresis. A total of 37 individuals participated in this study of which 12 and 25 of them were categorized in autologous and allogeneic groups respectively. For the apheresis procedure, a central venous access was used as well as the custom prime method with some changes. Apheresis details, as well as CD34 and CD3 cell counts in the allogeneic and autologous groups, were calculated. In this study, 91.9% (N = 34) of all individuals achieved the minimal amount of cells for PBSCT (2 × 106 CD34+ cells/kg) in one session. On the other hand, 12% (N = 3) of donors in the allogeneic group achieved the minimal threshold in 2 apheresis sessions. During the leukapheresis a total processed blood volume/total blood volume ratio (TPBV/TBV) was calculated as 4.64 ± 1.06 and 5.18 ± 0.73 fold in the allogeneic and autologous groups respectively. The mean of harvested CD34 cells in allogeneic and autologous groups was 5.28 ± 3.47 × 106 and 3.57 ± 2.9 × 106 cells/kg respectively. Likewise, in the allogeneic group, the mean of the harvested CD3 cell count was 339 ± 141 × 106/kg. Also, the median day of white blood cell (WBC) engraftment was 14 and 13 for allogeneic and autologous groups respectively. Furthermore, the median day of platelet engraftment was 19.5 for both allogeneic and autologous groups. Among the recipients of the allogeneic group, acute graft versus host disease (aGVHD) was detected in 56% (N = 14) of patients and this was also correct for chronic GVHD. Taken together, it was shown, despite the probable complications of peripheral blood stem cell apheresis in donors weighing less than 20 kg; that it is possible to perform this procedure without any complication during the leukapheresis.

Autologous Platelet-Released Growth Factor and Sexual Dysfunction Amendment: A Pilot Clinical Trial of Successful Improvement Sexual Dysfunction after Pelvic Irradiation

Sexual dysfunction (SDF) is a common sequel to cancer treatment which affects the quality of life in women treated with pelvic radiotherapy. The aim of this study was to evaluate the safety, symptom resolution and objective improvement the injection of autologous platelet released growth factor (APRGF) for treatment of SDF in cited patients. This prospective pilot study enrolled 10 cancer-free patients with SDF who underwent pelvic radiotherapy at least 5 years ago, randomly. Each patient was received 1-2 cc APRGF within four weeks and all patients were re-evaluated at eight weeks and six months. CD34 immuno histochemistry and Masson's trichrome staining were performed on vaginal biopsy section for angiogenesis and fibrosis assay respectively. Sexual satisfaction after the injection of APRFG was clinically difference and the entire patient had sexual satisfaction. In the patient's follow-up, none of them needs to repeat the treatment. Our results declared that APRGF injection was effective and symptoms were disappeared in the entire patients. Significant objective improvements in vaginal diameter (mean before injection, 6.5 cm vs 7.1 cm after injection) (p-value = 0.001) and vaginal flexibility (mean before treatment, 0.72 cm vs 1.85 cm after injection) (P-value = 0.026) were observed. Characteristics of discharge before the injection in 60% of patients were included dry vagina and 40% had mild discharge but after injection 40% of patients had moderate and also 60% had mild and sufficient discharge (P-value= 0.190). Overally, our patients reported better sexual function and showed better vaginal function indexes, after APRFG injection.

How do anxiety affect CD34 and CD3 cells in allogeneic peripheral blood stem cell transplantation?

Allogeneic peripheral blood stem cells (PBHSCs) transplantation using apheresis is a curative method for malignant and non-malignant hematologic diseases. The aim of this study was to assess the possible effects of anxiety as well as other variables on PBHSCs apheresis success. In this cross-sectional study, different demographic and clinical data such as granulocyte-colony stimulating factor (G-CSF) dosage, CD 34+ cells count (before apheresis), CD 34+, CD3+ cells count in apheresis product and also complete blood count were assessed. Furthermore, for evaluation of anxiety level in donors, the Beck's anxiety inventory (BAI) was administered. In this study, 111 donors were randomly enrolled after meeting inclusion and exclusion criteria. Results of BAI showed the mean score of 22.85 ±â€¯15.43 (mild to moderate anxiety) for the participants. According to the bootstrapped quantile regression analysis, a statistically significant linear association was found between CD34+ cell count and BAI score (P-value < 0.001) after adjusting for confounding variables. Moreover, the BAI score had a statistically significant effect on CD3+ cells count (P-value = 0.021) after adjusting for confounding variables. Taken together, results showed that anxiety affects both CD34+ and CD3+ cells count. Thus, the authors suggest that anxiety levels would be evaluated as well as other variables in donors in order to run a proper intervention by professionals.

Extra corporeal photochemotherapy in steroid refractory graft versus host disease: A review of guidelines and recommendations.

Regardless of remarkable progresses in prevention and treatment approaches, graft versus host disease (GVHD) remains a major impediment for successful allogeneic hematopoietic stem cells transplantation (HSCT) and leads to morbidity and mortality in transplanted patients. Corticosteroids are the standard therapy for GVHD; however, a great number of patients will not respond sufficiently and others will be significantly affected by adverse effects of steroids. Extracorporeal photochemotherapy (ECP), as one of the numerous second line therapies, through modulation of immune cells may improves GVHD affected organ function in steroid-refractory forms. Considering to widespread utilization of ECP as a therapeutic strategy, we performed review on current literature of ECP, regarding the treatment strategies, monitoring protocols and technical aspects in chronic and acute GVHD.

Optimizing Stem Cells Mobilization Strategies to Ameliorate Patient Outcomes: A Review of Guide- lines and Recommendations.

Peripheral blood stem cell transplantation (PBSCT) is an effective treatment for hematological malignancies. Mobilization of peripheral blood progenitor cells performs in different ways among transplantation centers. Forceful mobilization schedules are comprised of growth factor alone, chemotherapy along with growth factor and also, a newly combination of novel agent such as plerixafor with any approach. With the appearance of numerous modifications in stem cell mobilization field over the past decade and advent of novel stem cell mobilization techniques, it seems to be necessary to review recent publications about stem cell mobilization strategies to respond above cited issues. Relevant literature was identified by a PubMed search (1996-2016) of English-language literature using the terms mobilization, Allogeneic Stem Cells Transplantation, Autologous Stem Cells Transplantation and technical aspects of apheresis. Although many institutions have established their own procedures to improve stem cell mobilization success rates accompanying cost-effectiveness considerations, an optimal stem cell mobilization regimen and methods have not been well-defined, yet. Practical guidelines are required to address critical clinical issues including proper growth factor, the most Impressive chemotherapy and its dosage and appropriate time for leukapheresis initiation. Hence, based on literature, we prepared practical guidelines in this review.

Evaluation of platelet-rich plasma gel potential in acceleration of wound healing duration in patients underwent pilonidal sinus surgery: A randomized controlled parallel clinical trial.

OBJECTIVES: One of the most important surgical issues applied in the treatment of pilonidal sinus disease is wound healing. The aim of this study was to investigate the possible effect of platelet-rich plasma (PRP) gel on accelerating wound healing in these patients. METHODS: In this randomized, controlled, parallel group clinical trial, 110 patients were randomly allocated into two parallel groups with the same size (controls and treatment arm) after meeting inclusion and exclusion criteria. After the surgery, controls were treated by classic wound dressing while the case group was treated with PRP gel in a classic wound dressing platform. The patients were then evaluated for duration of antibiotics consumption, experienced pain and the time of returning to routine activities. Also, both groups were assessed for angiogenesis (by detecting CD34+ cells using immunohistochemical assay) and collagen sedimentation (masson's trichrome staining) using pre-complete healing wound biopsy. All the statistical analyses were performed using SPPS 20 and p-values of less than 0.05 considered statically significant. RESULTS: According to the results, patients treated with PRP gel went through a significantly faster healing process (8.69±1.18 in controls and 4.78±0.87 weeks in PRP gel treated ones with the P-value=0.03) and returned to their routine activities (3.3±0.64 for the treatment of arm and 6.5±1.03 weeks for controls with the P-value=0.00) while experiencing less pain (P-value=0.00) and shorter anti-biotic consumption duration (P-value=0.00). CONCLUSION: Considering the results, authors of this study suggest PRP gel treatment for post operation wound dressing of pilonidal sinus disease with healing by secondary intention.

Human Platelet Lysate as a Xeno Free Alternative of Fetal Bovine Serum for the In Vitro Expansion of Human Mesenchymal Stromal Cells.

BACKGROUND: Mesenchymal stromal cells (MSCs) are employed in various different clinical settings in order to modulate immune response. Human autologous and allogeneic supplements including platelet derivatives such as platelet lysate (PL), platelet-released factors (PRF) and serum are assessed in clinical studies to replace fetal bovine serum (FBS). The immunosuppressive activity and multi-potential characteristic of MSCs appear to be maintained when the cells are expanded in platelet derivatives. MATERIALS AND METHODS: Platelet-rich plasma was collected from umbrical cord blood (UCB). Platelet-derived growth factors obtained by freeze and thaw methods. CD62P expression was determined by flow cytometry. The concentration of PDGF-BB and PDGF-AB was detemined by ELISA. We tested the ability of a different concentration of PL-supplemented medium to support the ex vivo expansion of Wharton's jelly derived MSCs. We also investigated the biological/functional properties of expanded MSCs in presence of different concentration of PL. The conventional karyotyping was performed in order to study the chromosomal stability. The gene expression of Collagen I and II aggrecan and SOX-9 in the presence of different concentrations of PL was evaluated by Real-time PCR. RESULTS: We observed 5% and 10% PL, causing greater effects on proliferation of MSCs .These cells exhibited typical morphology, immunophenotype and differentiation capacity. The genetic stability of these derivative cells from Wharton's jelly was demonstrated by a normal karyotype. Furthermore, the results of Real-time PCR analysis showed that the expression of chondrocyte specific genes was higher in MSCs in the presence of 5% and 10% PL, compared with FBS supplement. CONCLUSIONS: We demonstrated that PL could be used as an alternative safe source of growth factors for expansion of MSCs and also maintained similar growing potential and phenotype without any effect on chromosomal stability.