Leukemia SH-1 cells purged by ZnPcH(1)-based photodynamic therapy / 中国实验血液学杂志

The objective of this study was to investigate the effect of a novel Zinc phthalocyanine (ZnPcH(1)) based photodynamic therapy (PDT) on acute monocytic leukemia cell lines SHI-1 and its mechanism, so as to provide theory basis for bone marrow purging in vitro for patients with leukemia. The killing effect of ZnPcH(1)-PDT on SHI-1 cells were assessed by MTT method; the SHI-1 cell death patterns were analyzed by AO/EB fluorescence staining, TdT-mediated dUTP nick end labeling (TUNEL), DNA ploidy analysis, and Annexin V-FITC/PI double staining.Cell mixture was established by integrating SHI-1 cells with normal bone marrow MNC (by 1:100-1:10 000). Purging effect of ZnPcH(1)-PDT against SHI-1 mixed into normal MNC was assessed by analyzing the expression of fusion gene MLL/AF6 mRNA using nested RT-PCR. The results showed that ZnPcH(1)-PDT could effectively inhibit SHI-1 cell proliferation in dose-dependent manner, and ZnPcH(1)-PDT could induce cell apoptosis in time-dependent manner. 0.5 µmol/L ZnPcH(1)-PDT could completely photoinactivated kill SHI-1 cells in the simulated remission bone marrow. It concluded that ZnPcH(1)-PDT may be a effective and convenient promising purging technique for leukemia.

Acute graft versus host disease in non-myeloablative allogeneic stem cell transplantation / 中国实验血液学杂志

The objective of this study was to explore the occurrence and clinical features of acute graft versus host disease (aGVHD) in non-myeloablative stem cell transplantation (NAST). 19 cases developed aGVHD out of 71 cases with NAST in recent years were analyzed retrospectively. Out of 19 cases, 9 males and 10 females at the median age of 38 (18-59), 16 cases with grade I-II aGVHD, 3 cases with grade III-IV aGVHD. The results indicated that the incidence of aGVHD in NAST was 26.7% (19/71), and severe aGVHD was 4.2%, the median onset time was 58 days (17-240 days) after transplantation. Skin and especially the intestine were the main target organs of aGVHD, while diarrhea occurred as the first symptom in 7 cases, 3 cases showed mixed acute and chronic GVHD involving more locations at the same time. aGVHD occurrence was 38.2% in those patients with full donor chimerism (FDC) and 16% in patients with the mixed chimerism (MC). It is concluded that aGVHD in NAST is less in occurrence, lighter in severity and later in time, but higher occurrence in those with early FDC, which intestine and skin are the main target organs. The clinical course is prolonged and easily complicated with severe infection in the later phase. Early combined therapy with powerful supportive treatment is necessary.

Establishment and application of a murine transplant model of bone marrow purging of metastatic breast cancer cells in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a murine transplant model for bone marrow purging of metastatic breast cancer and to explore the efficiency of Econazole (Ec) as a purging agent.</p><p><b>METHODS</b>Mixtures of TSA /Neo breast cancer cells and murine bone marrow cells were transplanted into lethally irradiated mice following purging with Econazole or saline in vitro. The recipient mice were monitored for hematopoietic engraftment, appearance of metastatic nodules in lungs and the overall survival.</p><p><b>RESULTS</b>All the mice receiving i.v. injection of TSA cells developed metastatic lung nodules. The hematological recovery was not delayed in mice transplanted with Ec purged bone marrow. More importantly, metastatic lung nodules were not seen in Ec treated group and the overall survival was improved.</p><p><b>CONCLUSION</b>The purged metastatic breast cancer cell bone marrow transplant model was easily established and reproducible. Ec could be used to purge the bone marrow grafts contaminated with breast cancer cells.</p>

Trasplante de células hematopoyéticas en mieloma múltiple / Hematopoietic stem cell transplantation in multiple myeloma

Multiple myeloma (MM) is the second most common hematologic malignancy, affecting approximately 14,000 new patients per year in the United States. For over four decades, the standard treatment for MM has been a regimen of melphalan combined with prednisone. Using this treatment modality, complete responses are rare, and 50% of patients have had disease that was resistant to chemotherapy. Attempts have been made to improve the outcome of MM by administering combinations of I. V. polichemotherapy, but these treatments are equivalent in terms of overall survival. High-dose therapy with peripheral blood stem cell support can be applied safely in these patients and achieves significantly higher complete remission rates as well as better event-free survival and overall survival. However, neither tumor-cell purging, positive selection, intensification of conditioning with additional chemotherapeutic agents, nor total body irradiation have been shown to improve outcome. The role of tandem transplantation with high-dose melphalan seems to be a good selection of treatment in hospitals having all resources. Future research will include the combination of the best remission-induction regimen with tandem transplants and maintenance treatments (thalidomide, idiotype or dendritic cell vaccination) that will sustain complete remission. Development of non-myeloablative allogeneic transplantation in order to exploit the graft-versus myeloma effect provides an alternative for patients who have a compatible donor. Combining all of these modalities with the new drugs developed few years ago (thalidomide, bortezomib, revlimid), we hope that MM will become a manageable chronic disease and perhaps a curable disease at least for 30% to 40% of the patients.

El mieloma múltiple (MM) es la segunda patología oncohematológica más frecuente. En Estados Unidos son diagnosticados anualmente 14,000 casos nuevos. En las últimas cuatro décadas el tratamiento estándar ha sido la combinación de melfalán y prednisona. Con este régimen raramente se logran remisiones completas y 50% de los pacientes no responden a esta terapia. Se han hecho intentos de mejorar los resultados combinando poliquimioterapia, pero la sobrevida global ha sido la misma. Al aplicar quimioterapia a dosis altas y rescate con trasplante de células hematopoyéticas se logra un mayor porcentaje de remisiones completas, asimismo, una mayor sobrevida libre de enfermedad y sobrevida global. La purga de células hematopoyéticas, selección positiva, intensificación del régimen de acondicionamiento con otras drogas o irradiación corporal total, no han demostrado utilidad en términos de sobrevida global. El doble trasplante autólogo de células hematopoyéticas parece ser una opción útil para hospitales que cuentan con la infraestructura y los recursos necesarios para realizarlo. En un futuro, la investigación deberá incluir el uso del mejor régimen de inducción a la remisión más doble trasplante autólogo y terapia de mantenimiento (talidomida o vacunas con células dendríticas), con la finalidad de al menos prolongar la remisión completa. El uso del trasplante alogénico no mieloablativo para provocar el efecto injerto contra mieloma parece una buena alternativa para los pacientes que tengan donador. Al combinar todas estas modalidades de tratamiento con las nuevas drogas desarrolladas en los últimos años (talidomida, bortezomid, revlimid), se espera que en un futuro el MM se convierta en una enfermedad crónica y curable en al menos 30 a 40% de los enfermos.

Experimental studies of the effects of ZnPcS2P2-based-photodynamic therapy on bone marrow purging / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>An effective purging technique plays an important role in autologous hematopoietic stem cells transplantation. Photodynamic therapy (PDT) provides a novel approach for this purpose. This study dealt with the purging effects of di-sulfo-di-phthalimidomethyl phthalolcyanine zinc (ZnPcS2P2)-based photodynamic therapy (ZnPc-PDT).</p><p><b>METHODS</b>Fluorescence intensity of cell extracts was measured using a fluorescence spectrophotometry. The proliferative potency of K562 cells and HL60 cells was detected using MTT colorimetric assay, Typan blue dye exclusion method, colony formation test. The proliferative potency of normal hematopoietic cells was evaluated using mixture colony-forming unit (CFU-Mix), granulocyte-macrophage colony-forming unit (CFU-GM), and erythrocyte colony-forming unit (CFU-E) assays. K562 cells were mixed with normal mononuclear cells (MNCs) at ratios of 1:100 and 1:1000 for creating the model of simulated remission bone marrow. Colony formation test and nested-RT-PCR were carried out to detect the residual K562 cells in cell mixture.</p><p><b>RESULTS</b>After a 5-hour incubation with ZnPcS2P2, the content of ZnPcS2P2 in normal MNCs was the lowest value. At the same time, the content in K562 cells and HL60 cells was very high. Therefore, the time point was selected as the optimal one for irradiating the cell suspensions. ZnPc-PDT could significantly kill proliferative K562 cells and HL60 cells in a dose-dependent manner. At the concentration of 1.0 microg/ml, the inhibitory rate of ZnPc-PDT on the colony formation was 100% for K562 cells, 89.7% for HL60 cells. 0.25 microg/ml ZnPc-PDT could completely photoinactivate residual K562 cells in the simulated remission bone marrow. Under an identical condition, the inhibitory rates of CFU-Mix, CFU-GM, CFU-E were 18.0%, 18.6%, and 17.8% respectively.</p><p><b>CONCLUSION</b>ZnPc-PDT appears to be a promising approach for bone marrow purging.</p>

Cistitis hemorrágica relacionada al trasplante de médula ósea: primer reporte de casos en el país / Hemorrhagic cystitis in bone marrow trasplant recipients: first report of cases in our country

Objetivo: Determinar la incidencia, características clínicas y complicaciones de la cistitis hemorrágica (CH). Material y Métodos: Estudio descriptivo sobre pacientes transplantados de médula ósea en la Unidad de Trasplante de Médula Ósea del Hospital Nacional Edgardo Rebagliati Martins, desde noviembre 1994 hasta octubre 2003, y que desarrollaron cistitis hemorrágica. Resultados: En 9 años fueron transplantados 170 pacientes, desarrollando cistitis hemorrágica 8 de ellos (incidencia acumulada 4,7 por ciento). El cuadro clínico se caracterizó por disuria más hematuria macroscópica o microscópica. La presentación tardía fue la más frecuente. En 4 pacientes, la cistitis hemorrágica fue severa. Dos de los tres pacientes que desarrollaron el cuadro de cistitis hemorrágica en dos oportunidades, fallecieron. Conclusiones: La mitad de los pocos pacientes que desarrollan CH en la Unidad, lo hacen bajo la forma severa y, si presentan la CH por segunda vez, su pronóstico se complica.

Objective: To determine the incidence, clinical features and complications of bone marrow transplant-related hemorrhagic cystitis (HC). Material and Methods: Retrospective study of bone marrow transplant patients attended at the Edgardo Rebagliati Martins National Hospital Bone Marrow Transplant Unit from November 1994 to October 2003 and who developed HC. Results: A total of 170 patients underwent hematopoietic cell transplantation. Eight patients developed HC with a cumulative incidence of 4,7 per cent, characterized by mainly late onset hematuria and dysuria. In 4 patients, the HC was severe. Two of the three patients who developed HC twice died. Conclusions: Half of the few patients with HC attended at the Unit developed serious disease and when they had HC for a second time the prognosis was fatal.

Comparison of autologous and allogeneic hematopoietic stem cell transplantation for 140 patients with de novo acute leukemia in first complete remission / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the outcome of patients with de novo acute leukemia (AL, no AML-M(3)) in CR(1) undergone autologous hematopoietic stem cell transplantation (auto-HSCT) or HLA-identical sibling allogeneic HSCT (allo-HSCT).</p><p><b>METHODS</b>Forty-six AL patients received allo-HSCT and 94 received auto-HSCT in CR(1). The conditioning regimens mainly consisted of TBICy, BuCy and MAC. Cyclosporine plus methotrexate, or cyclosporine alone, or FK506 alone was used for graft-versus-host disease (GVHD) prophylaxis. Among auto-HSCT group, 39 patients received purged autologous bone marrow and 38 received immunotherapy and/or maintenance chemotherapy after transplant.</p><p><b>RESULTS</b>Myeloid reconstitution was achieved in all patients. After a median of 700 (range, 18 approximately 5563) days follow-up, the probabilities of leukemia-free survival (LFS) at 5 year were not significantly different in these two groups: (51.5 +/- 5.4)% for auto-HSCT group and (52.8 +/- 7.6)% for allo-HSCT group (P > 0.05). There was a lower cumulative relapse incidence (RI) [(26.3 +/- 6.9)% vs. (52.0 +/- 5.5)%, P > 0.05] but a significantly higher cumulative transplant-related mortality (TRM) [(37.6 +/- 7.8% vs. (14.4 +/- 4.1)%, P < 0.05] in the allo-HSCT group than in auto-HSCT group. Among auto-HSCT group, the patients received purged autografts and/or post-transplant therapy had significantly better LFS and lower RI (P < 0.05) than those received unpurged autografts or no post-transplant treatments [5-y LFS: (62.8 +/- 6.8)% and (38.4 +/- 8.4)%; RI: (37.7 +/- 6.8)% and (74.2 +/- 8.7)%, respectively].</p><p><b>CONCLUSION</b>The long-term LFS of auto-HSCT was comparable to that of allo-HSCT in the management of patients with AL in CR(1), because autograft purging and post-transplant treatment can significantly decrease relapse of auto-HSCT patients and auto-HSCT has lower therapy-related toxicities.</p>

Myeloablative Treatment Supported by Autologous Stem Cell Infusion with Neuroblastoma

Bcr-abl antisense oligodeoxynucleotides (AS-ODNs) have provided evidence of an antileukemia effect when tested in vitro against Philadelphia-positive cells. In order to investigate the efficacy of AS-ODNs as purging agents in chronic myeloid leukemia (CML) patients, K562 cells, a human CML cell line, were treated in vitro with various types of AS-ODNs and interferon-alpha. Cells were treated in vitro for 0 and 36 hr with 40 microgram/mL of AS-ODNs, respectively, and incubated at 37 degrees C for 36 hr. Cytotoxic effects were measured by counting the number of viable cells as well as by MTT test. Clonogenic activities were evaluated by methylcellulose culture for 2 weeks. The effects of purging agents on the rearrangement of bcrabl gene were evaluated by RT-PCR. AS-ODNs inhibited the proliferation of K562 cells with time in cell count assay and MTT test. AS-ODNs were superior to INF-alpha in inhibiting clonogenic activity (recovery rate; 26.3% vs 64.0%). After incubation with bcr-abl AS-ODNs primers and mRNA isolated from K562 cells, positive bands were abolished, especially of b3a2 type and phosphorothioate type. Our results suggest that AS-ODNs mediated purging may be one of the efficient methods and that autograft may be an alternative treatment for allograft in high-risk group patients of CML if they do not have a stem cell donor.

Comparison of curative effect of autologous peripheral blood stem cell transplantation versus bone marrow transplantation for acute leukemia / 中国实验血液学杂志

To compare the clinical outcome of autologous peripheral blood stem cell transplantation (APBSCT) and autologous bone marrow transplantation (ABMT) in treatment of patients with acute leukemia in first remission, 41 patients received APBSCT, 17 patients received unpurged ABMT and 30 patients received purged ABMT. The results showed that hematopoietic recovery was significantly earlier after APBSCT than that after purged or unpurged ABMT. The 3-year disease-free survival (DFS), relapse rate (RR) and transplant-related mortality (TRM) for all patients of 3 groups were 51.7%, 41.7% and 6.8%, respectively. DFS and RR were significantly influenced by disease types (ALL or AML) and intervals between diagnosis and CR(1) or CR(1) and transplant. The main causes of transplant-related death were infection and hemorrhage. After APBSCT, DFS, RR and TRM were 48.4%, 43.9% and 4.9%, respectively, and did not differ significantly from those found in unpurged ABMT (47.1%, 45.6% and 11.8%) or purged ABMT (66.5%, 29.6% and 6.7%). It is concluded that the clinical outcome of APBSCT is similar to unpurged or purged ABMT but APBSCT allows faster recovery of hematopoiesis and needs less transfusion support.

FasL-cDNA transfected into mouse bone marrow cells ex vivo to prevent graft versus host disease / 中国实验血液学杂志

To explore the new approach to prevent graft versus host disease (GVHD) by purging ex vivo T lymphocytes of bone marrow graft through Fas-FasL way, FasL-cDNA was transfected into BALB/c mouse bon e marrow cells by liposome ex vivo. The transfected cells were cultured together with BAC (BALB/c x C57BL/6) mouse bone marrow graft. The mixing bone marrow graft was infused into BALB/c mouse recipients after 60Co-gamma irradiation. The mortality, manifestation and pathologic change of GVHD in recipient mice were observed. The CFU-S and Y chromosome from donor mice were detected. The results showed that compared with control group, the mortality in 60 days of the recipients in the experimental group decreased (20% vs 70%, P < 0.01) and the morbidity of GVHD lowered (40% vs 100%, P < 0.01). The CFU-S counts for all groups were at normal level on 20 days after transplantation. The Y chromosome from donor mice was discovered in 70% bone marrow nucleated cells of recipient mice survived over 2 months in the experimental group. It is concluded that mFasL-cDNA transfected mouse bone marrow cells prevent GVHD after culturing together with bone marrow graft, and accelerate hematopoietic reconstitution in recipient mice.

Establishment of models for purging leukemic cells from the grafts in vitro / 中国实验血液学杂志

Using a fluorochrome Calcein-AM, leukemia cells were labeled and seeded into cell lines or bone marrow cells to establish three cell-models of grafts with leukemia. These cell-models were engaged with CD34 immunomagnetic beads and the purging efficacy was evaluated using both fluorescence microscopy and flow cytometry. The results showed that the cell-models established in this study could be evaluated successfully not only with fluorescence microscopy but also flow cytometry. After CD34 positive selection, KG1a cells were removed by (0.98 +/- 0.09) log in model II and NALM-6 cells were removed by (1.82 +/- 0.51) log in model III, respectively. It is concluded that the models established in this study are stable and direct with an excellent reproducibility and an accuracy, which can be used to evaluate purging efficacy of leukemia cells in model graft using immunomagnetic selection and the experimental studies on tumorcidal effect in vitro.

Síndrome hemofogocítico asociado a citomegalovirus después de trasplante de médula ósea autólogo y purga con mafosfamida / SCitomegalovirus-associated hemophagocytic syndrome after auatologous bone marrow transplantation purged with maphosphamide

Introducción. El síndrome hemofagocítico (SH) es una proliferación benigna de histiocitos, asociado con fagocitosis de elementos hemopoyéticos. El cuadro clínico se caracteriza por: fiebre, linfadenopatía, hepatoesplenomegalia, citopenias profundas y coagulopatías. Se ha asociado a: virus, bacterias, hongos, parásitos y neoplasias. Caso clínico. Se informa el caso de un SH después de trasplante de médula ósea autólogo y purga con mafosfamida en un paciente con leucemia aguda mieloblástica M-4 sin donador HLA compatible, en primera remisión completa con serología y reacción en cadena de la polimerasa (PCR) para citomegalovirus negativos (CMV) antes del trasplante, todas sus transfusiones se aplicaron con filtros leuco-reductores. La toma de injerto fue en el día +17. En el día +28 presentó neutropenia de 200 /mL, el aspirado de médula ósea mostró hemofagocitosis y PCR para CMV fue positiva, en leucocitos, plasma y orina; después de tratamiento con ganciclovir e inmunoglobulina intravenosa la PCR se negativizó y la médula ósea se observó sin evidencia de hemofagocitosis; no se documentaron otras infecciones o neoplasias.Conclusión. De acuerdo a las características clínicas y estudios de laboratorio, se consideró que la hemofagocitosis intramedular estuvo asociada a la infección por CMV.

Therapeutic apheresis.

During the last 30 years in vivo blood cell separation, generally referred to apheresis, has established a central role in both blood donor programmes and therapeutics. The technological advances in apheresis equipment has made procedures safer, faster and more effective. This article will review the use of apheresis in clinical medicine with emphasis on plasma exchange and peripheral blood stem cell collection. Plasma exchange now has a pivotal role in the management of a range of disorders, specially those with autoimmune pathogenesis. However, Plasma exchange should be practised as one component of an integrated and frequently multidisciplinary approach to management. The harvesting of allogeneic or autologous of peripheral blood haemopoietic stem cells is increased and it has become the principle indication for apheresis in many haematology units. A well coordinated protocol approach to this procedure is important if adequate haemopoietic stems cells are to be collected and safely cyropreserved. This requires successful cooperation between medical, nursing and scientific personnel.

Results of allogeneic bone marrow transplant in chronic myeloid leukaemia following conditioning with busulfan and cyclophosphamide.

OBJECTIVE: To study the outcome of oral busulfan and intravenous cyclophosphamide (BuCY 2 regimen) followed by allogeneic bone marrow transplant (BMT) in a cohort of patients with Philadelphia chromosome (Ph+) chronic myeloid leukaemia (CML) in a single centre. METHODS: From 1991 to March 1998, a total of 27 consecutive Ph+ CML patients received busulfan 4 mg/kg/day over 4 days and cyclophosphamide 60 mg/kg/day over 2 days followed by infusion of HLA-identical sibling haematopoietic stem cells. All except one (who received peripheral blood stem cells) were given donor bone marrow cells. Post-transplant graft versus host disease (GVHD) prophylaxis included a short course of methotrexate (on days +1, +3, +6 and +11) and cyclosporine till day +180. RESULTS: With a median follow-up of 30.5 months (1-55+ months), 14 patients (52%) are alive free from relapse. Early mortality was relatively high with 10 patients (37%) dying within first 100 days post-transplant. Acute GVHD developed in 14 patients (52%) inspite of GVHD prophylaxis with methotrexate and cyclosporine; six had grade I/II and eight grade III/IV. Chronic GVHD developed in five of 15 patients who lived beyond 70 days. CONCLUSION: Allogeneic BMT appears to result in eradication of CML and ensure disease free survival in about half of the young patients. However, efforts should be on to minimise early mortality.

Inhibition of Overexpression of c-myb Protooncogene in K562 Cells Using c-myb Antisense Oligodeoxynucleotides / 대한소아혈액종양학회지

PURPOSE: The c-myb protooncogene encodes MYB protein that is critical for normal and leukemic hematopoietic cell proliferation and development. It is known that c-myb plays an important role in leukemogenesis as well. Aberrant expression of c-myb is seen in acute myeloid leukemia (AML), acute lymphoid leukemia (ALL), and chronic myeloid leukemia (CML). We reasoned that down regulation of c-myb expression using synthetic antisense oligomers targeted to c-myb mRNA might prove useful antileukemic agents if leukemia cells were more dependent on c-myb function for proliferation than their normal counterparts. To investigate the applying possibility of c-myb antisense oligodeoxynucleotides as the useful antileukemic agents, we examined the cell viability, cloning efficiency and the expression of c-myb mRNA and MYB protein after the exposure of c-myb oligomers on normal bone marrow cells and leukemia cells. MATERIALS AND METHODS: We maintained in short-term suspension culture for 4 days to analyze the effect of c-myb oligomers on normal bone marrow cells and chronic myelocytic leukemia cell line K562. During suspension culture, cell counts and viability were periodically determined, and immediately seeded into duplicate methylcellulose cultures containing recombinanat human interleukin 3 and GM-CSF. Cells placed in semisolid cultures were allowed to grow for an additional 10~12 days. We counted the colonies, and then RNA and protein was extracted from cells cloned in liquified methyl- cellulose cultures. We detected the c-myb mRNA expression by RT-PCR and Southern hybridization analysis and MYB expression by Western hybridization analysis. RESULTS: c-myb sense oligomers had negligible effects on K562 cells growth in short- term suspension culture, while exposure to c-myb antisense oligomers resulted in a daily decline in cell number. In contrast, normal bone marow cell viability and numbers were unaffected by c-myb sense or antisense oligomers exposure. c-myb antisense oligodeoxy nucleotides strongly inhibited or completely abolished growth and colony formation of K562 cells. In contrast, c-myb sense oligomers did not affect. At antisense dose that inhibited leukemic cell growth, normal bone marrow cells survived. Thus, normal and leukemic cells showed the differential sensitivity to the toxic effect of c-myb antisense oligomers. RT-PCR, Southern hybridization analysis and Western hybridization analysis of c-myb antisense-treated K562 cells revealed a complete absence of c-myb mRNA expression and MYB expression. CONCLUSION: Results obtained from these studies suggest that inhibition of c-myb function with antisense oligodeoxynucleotides might eventually form the basis for a molecular biologic approach to leukemia therapy, perhaps most immediately as ex vivo bone marrow purging agent.

Evaluating efficiency of bone marrow harvest and its manipulation--role of CD 34 positivity.

We evaluated harvested marrow cells for total nucleated cells (27.49 x 10(9)), absolute 'lymphocyte' count (6.29 x 10(9)) and CD 34 positive cells (3.57 x 10(9)). The same parameters were studied after in vitro manipulation to remove RBCs and plasma. Reinfused WBCs contained 12.87 x 10(9) nucleated cells, 4.25 x 10(9) absolute 'lymphocyutes' and 3.34 x 10(9) CD 34 positive cells. The corresponding figures for loss during in vitro manipulation (tubing, RBCs and plasma) are 14.62 (53.18%), 2.04 (32.43%) and 0.23 x 10(9) (6.44%) cells respectively. Therefore CD 34 positivity may be a better indicator of total yield, loss during manipulation and reinfusion of hemopoietic progenitor cells in bone marrow transplantation.