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At least 775 records · Page 43Linked to original sources
PubMed1993

Protection of bone marrow stromal cells from the toxic effects of cyclophosphamide in vivo and of ASTA-Z 7557 and etoposide in vitro by ammonium trichloro(dioxyethylene-O-O')tellurate (AS101).

The immunomodulator AS101 has previously been shown to protect mice from lethal and sublethal doses of cyclophosphamide (CYP). AS101 was also shown to protect BM granulocyte-macrophage colony-forming cells from the toxic effects of ASTA-Z 7557. In the present study we examined the ability of AS101 to protect functional properties of BM stromal cells from the toxic effects of CYP in vivo or ASTA-Z in vitro. The functional properties of stromal cells from CYP-injected mice were tested with respect to stromal cell number and viability as reflected by the number of colony-forming unit fibroblasts, the ability of established stromal layers to secrete colony-stimulating factor and interleukin 6, as well as the capacity to support hemopoietic cells. All of these parameters were tested from day 1 to day 7 after CYP treatment. We demonstrate that all stromal functions are severely damaged following CYP treatment. Pretreatment of mice with 10 micrograms AS101 24 h before injection of 250 mg/kg CYP resulted in a significant amelioration of stromal cell functions as early as 24 h following CYP treatment. In addition we show that prior incubation of BM cells with AS101 protects the development of stromal colony-forming unit fibroblasts from the toxic effects of ASTA-Z, a potent derivative of CYP, and etoposide, a derivative of podophyllotoxin. These results strongly suggest the usefulness of AS101 in counteracting chemotherapy-induced BM microenvironmental suppression and the important role of the compound as an adjunct treatment of cancer when used in combination with CYP. The data also suggest the effectiveness of AS101 in purging bone marrow when used concomitantly with ASTA-Z or etoposide.

Adjuvants, Immunologic↗
PubMed1995

Immunotherapy of multiple myeloma.

In 1994, an estimated 12,700 new cases of multiple myeloma (MM) will be diagnosed in the USA and 9,800 patients will die from this disease. At present, a cure for MM has not been achieved with any chemotherapeutic regimen. Therefore, it is important to develop novel therapeutic approaches to treat this fatal disease. This review focuses on new concepts in the immunotherapy of MM. Thus far, interferons and anti-human interleukin (IL)-6 monoclonal anti-bodies (MAbs) have been used to treat patients with this disease. Bone marrow transplantation using autologous marrow purged with MAbs and complement, with anti-myeloma immunotoxins (ITs), or MAb-magnetic bead conjugates has been reported. Adoptive cellular therapy, in vivo with anti-CD3 and IL-2, as well as transplantation of purified autologous CD34+ peripheral blood stem cells, is now being evaluated in clinical trials. Anti-human IL-6 receptor (IL-6R) and anti-CD54 (ICAM-1) MAbs have shown promising results in the therapy of human myeloma cell lines in SCID mice, while an IL-6 antagonist protein, anti-gp130 MAbs, recombinant soluble gp130, anti-B7, anti-HLA-DR, and recombinant soluble CD16 also inhibit the growth of myeloma cell lines in vitro. These experimental therapeutic modalities hold promise for use in humans and may also provide further insights into the pathogenesis of MM.

Antibodies, Monoclonal↗
PubMed1993

Autologous bone marrow transplantation for acute myeloid leukemia using busulfan plus etoposide as a preparative regimen.

We have studied the use of a new preparative regimen for the treatment of patients in remission of acute myeloid leukemia (AML) with autologous bone marrow transplantation. Chemotherapy consisted of busulfan 1 mg/kg every 6 hours for 4 days (total dose, 16 mg/kg) on days -7 through -4 followed by an intravenous infusion over 6 to 10 hours of etoposide 60 mg/kg on day -3. Autologous bone marrow, treated in vitro with 100 micrograms/mL of 4-hydroperoxycyclophosphamide, was infused on day 0. We have treated 58 patients up to the age of 60 years, 32 in first remission, 21 in second or third remission, and 5 with primary refractory AML unresponsive to high-dose Ara-C, but achieving remission with aggressive salvage regimens. Of the first remission patients, there has been 1 treatment related death and 5 relapses. With median follow-up of 22 months, the actuarial relapse rate is 22% +/- 9% and disease-free survival is 76% +/- 9% at 3 years. Patients with favorable French-American-British (FAB) subtypes (M3 or M4 EO) did especially well, with no relapses seen in 15 patients observed for a median of 30 months. Actuarial relapse rate at 3 years was 48% for first remission patients with less favorable FAB subtypes. Of patients in second or third remission, there were 5 treatment related deaths and 4 relapses. With median follow-up of 22 months, the actuarial relapse rate is 25% +/- 11% and disease-free survival is 56% +/- 11% at 3 years. Four of five primary refractory patients died during treatment and 1 remains in remission with short follow-up. These preliminary data are very encouraging and, if confirmed, support the use of autologous purged bone marrow transplantation using aggressive preparative regimens as one approach to improve the outcome of adults with AML.

Actuarial Analysis↗
PubMed2001

Improved intracellular delivery of oligonucleotides by square wave electroporation.

Prior studies have shown that electroporation is a simple and effective method for the introduction of oligonucleotides (ODN) into cells. In ex vivo bone marrow purging models, electroporation of ODN into cells has been associated with selective killing of human neoplastic cells while sparing hematopoietic stem cells. Prior studies used conventional electroporation methods (i.e., exponential decay) to introduce ODN into cells. Square wave electroporation allows the delivery of a more defined and regulated electrical pulse and is associated with high transfection efficiencies in a variety of systems. The current study was undertaken to determine whether square wave electroporation was more effective than exponential decay electroporation for the delivery of ODN into hematopoietic cells. Using fluorescein-tagged ODN and K562, chronic myelogenous leukemia (CML) cells, higher transfection rates were observed after square wave electroporation. In addition, c-myc antisense ODN were more effective in reducing c-myc protein when introduced by square wave electroporation, as compared with introduction by exponential decay electroporation. Square wave electroporation is thus identified as the optimal method for delivering ODN into hematopoietic cells.

Base Sequence↗
PubMed1990

The role of autologous bone marrow transplantation in 46 adult patients with non-Hodgkin's lymphomas.

Forty-six patients with non-Hodgkin's lymphoma (NHL) were treated with autologous bone marrow transplantation (ABMT) in two different institutions. All patients were pretreated with conventional chemotherapy. Three different conditioning regimens were used, and 20 patients underwent bone marrow purging. Twelve patients were treated in first complete remission (CR); eight are in unmaintained CR 8 to 104 months after ABMT. Five patients were grafted in first partial remission (PR) after conventional therapy; all achieved CR, and all remain in prolonged CR (first CR for four patients, second CR for one patient). Of 21 patients with chemosensitive relapses, 13 patients are in prolonged unmaintained CR 8 to 94 months after ABMT. Eight patients with resistant disease remained uncured by ABMT; all eight died, six from progressive illness and two from toxicity. The current 3-year disease-free probability is 60% for all patients, 0% for refractory disease; 82% for first PR or CR, and 60% for sensitive relapses (SRs). These results confirm the efficacy of ABMT in the treatment of chemosensitive NHL with bad prognosis.

Adolescent↗
PubMed1990

In vitro evaluation of combination drug purging for autologous bone marrow transplantation.

Using an in vitro model, we studied whether combining 4-hydroperoxycyclophosphamide (4HC) with other drugs could improve its effectiveness as an ex vivo purging agent for autologous bone marrow transplantation. 4HC was incubated simultaneously with vincristine and etoposide, and sequentially with methylprednisolone, in various combinations. Compared to 4HC alone, all drug combinations increased the kill of the leukemia cell lines K562 and CEM without increasing the kill of granulocyte-macrophage colony-forming units (CFU-GM). The combination of 4HC, vincristine and methylprednisolone was the most active, and this drug combination was also the only combination which showed improved selective cytotoxicity (compared to 4HC alone) toward REH cells. This combination inhibited at least 8 logs of clonogenic leukemia cells from all three cell lines at doses which spared 1% of CFU-GM. This was an increase of 1.7 to 6.6 logs of clonogenic leukemia cell kill over 4HC alone. This drug combination displayed similar differential activity between fresh clonogenic leukemia cells and CFU-GM cultured from the bone marrows of seven patients about to undergo autologous bone marrow transplantation for acute lymphocytic leukemia.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2005

Increasing incidence of late second malignancies after conditioning with cyclophosphamide and total-body irradiation and autologous bone marrow transplantation for non-Hodgkin's lymphoma.

PURPOSE: Although the risk of myelodysplastic syndrome (MDS) has been well-described following autologous bone marrow transplantation (ABMT), the risk of solid tumors has been poorly characterized. We report the incidence and outcome of solid tumors at 10-year follow-up in a large cohort of uniformly treated patients who underwent ABMT for non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Between 1982 and 1997, 605 patients underwent ABMT for B-cell NHL, with uniform conditioning with cyclophosphamide and total-body irradiation followed by reinfusion of autologous bone marrow purged with anti-B-cell monoclonal antibodies. Current information on relapse of disease and second malignancies was obtained via an institutional review board-approved questionnaire sent to the referring oncologists. RESULTS: Forty-two solid tumors, six non-MDS hematologic malignancies, 39 nonmelanoma skin cancers, and 68 cases of MDS/acute myelogenous leukemia (AML) were observed at a median follow-up of 9.5 years. A cumulative incidence model using death as a competing risk found that the 10-year incidence of second malignancy is 21%, with 10.0% non-MDS malignancies. The projected incidence of all malignancies at 15 years is 29%. The principal risk factor for second malignancy is increased age at ABMT (P = .0002). In the entire cohort, 9.6% of patients have died of second malignancy. CONCLUSION: Lengthy follow-up demonstrates a significant incidence of second malignancies after ABMT for NHL. Although the incidence of MDS/AML starts to plateau, the incidence of solid tumors continues to rise. Second malignancies are responsible for a significant fraction of overall mortality following ABMT.

Adult↗
PubMed2004

[Therapeutic efficacy of hematopoietic stem cell transplantation in patients with chronic myelogenous leukemia].

OBJECTIVE: To evaluate the clineffects of autologous and allogeneic hematopoietic stem cell transplantation (HSCT) for chronic myelogenous leukemia (CML). METHODS: Fifty-seven patients with CML were treated by HSCT, including 8 patients treated with autologous transplantation in vivo and vitro purging minimal residual disease (MRD), 39 with related donor allogeneic HSCT (allo-HSCT), and 10 with unrelated donor allo-HSCT. For the conditioning regimen, total-body irradiation with cyclophosphamide (CTX) was given in 32 patients, modified BuCY protocol (hydroxyurea, busulfan, Ara-C, CTX) in 24 patients, and MACC protocol (melphalan, Ara-C, CTX and lomustine) in one patient. Cyclosporine (CsA) and methotrexate (MTX) were used in patients with related donor allo-HSCT, and the combination of CsA, MTX , mycophenolate mofetil (MMF), and antithymocyte globulin (ATG) in unrelated donor all-HSCT to prevent graft versus host disease (GVHD). Kaplan-Meier survival analysis model was used to estimate the overall survival and the disease-free survival (DFS) at 5 years after transplantation. RESULTS: In 8 patients with autologous transplantation, 7 obtained partial or complete cytogenetic remission (CR) within 3 months after transplantation and one died of transplantation-related complication. In 49 patients with allo-HSCT transplantation, all patients obtained CR except for two patients, one of whom failed to obtain CR and the other died of hepatic veno-occlusive disease. The incidence of infection and veno-occlusive disease during transplantation was 33.3% and 7.0%, respectively. The incidence of hemorrhagic cystitis and cytomegalovirus interstitial pneumonia after transplantation was 22.8% and 8.8%, respectively. Veno-occlusive disease, hemorrhagic cystitis or cytomegalovirus interstitial pneumonia did not occur in patients with autologous transplantation. The incidence of acute GVHD was 41.0% and 48.6%, and that of chronic GVHD 40.0% and 42.9% in patients with related and unrelated transplantation, respectively. The rate of relapse was 57.1% and 12.8%, with DFS at 5 years of 25.0% and 61.7%, respectively, in patients with autologous and related donor transplantation. The DFS at 5 years was 70.7% and 34.1%, respectively, in patients with chronic/accelerated phases and blast crisis be fore trans plantation. CONCLUSION: allo-HSCT can produce a higher clinical cure rate in CML patients in chronic phase CsA+MTX+MMF+ ATG protocol is more effective for prevention and alleviation of acute GVHD in patients with unrelated donor transplantation. Autologous transplantation with bone marrow purging helps prolong the patients' survival and even obtain clinical cure of CML.

Adolescent↗
PubMed2004

[Hematopoietic stem cell transplantation for patients with chronic myelogenous leukemia].

BACKGROUND & OBJECTIVE: Chronic myelogenous leukemia (CML) is a malignant hematological disease.CML patients are commonly treated with hematopoietic stem cells transplantation (HSCT). This study was designed to evaluate the effect of HSCT on the patients with CML. METHODS: Forty-four patients with CML were treated by HSCT, including 8 cases treated with purging autologous transplantation, 30 cases with related donor allogeneic HSCT (allo-HSCT), and 6 cases with unrelated donor allo-HSCT. The conditioning regimen was TBI (total-body irradiation)+CY (CTX) protocol in 31 patients and modified BuCY (hydroxyurea, busulfan, Ara-C, CTX) protocal in 12 patients, and MACC (Melphalan, Ara-C, CTX and CCNU) protocol in one patient. CsA (cyclosporine) and MTX were used in the patients with related donor allo-HSCT, and CsA and MTX added to mycophenolate mofetil (MMF) and antithymocyte globulin (ATG) were used in the patients with unrelated donor all-HSCT for graft versus host disease (GVHD) prophylaxis. Otherwise, CsA was only used in the patients with accelerated phase(AP) and blast crisis(BC) for GVHD prophylaxis. Kaplan-Meier survival analysis model was used to estimate the overall survival (OS) and the disease-free survival (DSF) at 5 years after transplantation. RESULTS: Eight patients with autologous transplantation, except one case died of transplantation-related-complication, obtained part or complete cytogenetic remission within 3 months after transplantation. One patient, who was BC and obtained complete remission (CR) in hematology before transplantation,obtained complete molecular remission for 81 months after autologous transplantation. All patients obtained CR, except one patient died of hepatic veno-occlusive disease (VOD) and one case did not obtained CR, in 36 patients with allo-HSCT after transplantation. The incidence of infection and VOD during transplantation were 38.6% and 9.1%, respectively. The incidences of hemorrhagic cystitis (HC) and cytomegalovirus (CMV) pneumonia after transplantation were 15.9% and 11.4%, respectively. VOD, HC, and CMV pneumonia did not occur in the patients with autologous transplantation. The incidence of acute GVHD in the patients with related and unrelated donor transplantation were 40.0% and 33.3%, respectively. The incidence of chronic GVHD was 43.4% in the patients with related donor transplantation. The rates of transplant-related mortality (TRM) in the patients with autologous and allogeneic transplantation were 12.5% and 16.7%, respectively. The rates of relapse in patients with autologous and related donor transplantation were 37.5% and 13.3%, respectively. The DFSs at 5 years in patients with autologous and related donor transplantation after transplantation were 18.7% and 53.7%, respectively. The DFS at 5 years in patients with CP (chronic phase) or AP and BC before transplantation were 66.4% and 26.7%, respectively. CONCLUSION: all-HSCT shows higher clinical cure rate to CML patients with CP. CsA+MTX+MMF+ATG protocol is more effective for acute GVHD prophylaxis and can decrease the incidence and severity of acute GVHD in patients with unrelated donor transplantation. Autologous transplantation with bone marrow purged can prolong the survival time and a few patients may be cured with autologous transplantation in CML.

Adolescent↗
PubMed2006

Comparison of two immunomagnetic separation technologies to deplete T cells from human blood samples.

The objective of this study was to compare the performance of two immunomagnetic separation technologies to deplete T cells from buffy coats of human blood. Specifically, two versions of the commercial MACS(R) Technology: MiniMACS and SuperMACS, and a prototype, flow-through system, the QMS, were evaluated. Peripheral blood mononuclear leukocytes (PBL) were isolated from buffy coats and an immunomagnetic separation of CD3(+) cells was conducted using company and optimized labeling protocols. To mimic peripheral blood containing bone marrow purged hematopoietic stem cells, HSC, CD34 expressing-cells (KG1a) were spiked into PBL prior to T-cell depletion once optimized depletion conditions were determined. Once the labeling protocol was optimized, the MiniMACS system performed well by producing a highly enriched CD3(+) fraction, and a respectable level of depletion of T cells and recovery of KG1a cells in the depleted fraction; an average log(10) depletion of T cells of 2.88 +/- 0.17 and an average recovery of the KG1a cells of 60.8 +/- 5.94% (n = 14). The performance of the SuperMACS system was very similar with an average log(10) depletion of T cells of 2.89 +/- 0.22 and an average recovery of KG1a of 63.1 +/- 8.55% (n = 10). In contrast, the QMS system produced an average log(10) depletion of T cells of 3.98 +/- 0.33 (n = 16) with a corresponding average recovery of 57.9 +/- 16.6% of the spiked CD34+ cells. The aforementioned QMS performance values were obtained using sorting speeds ranging from 2.5 x 10(4) to 1.7 x 10(5) cells per second. It is suggested that the lack of a 100% recovery of the unlabeled KG1a cells is the result of a previously reported "drafting" phenomena which pulls unlabeled cells in the direction of the magnetically labeled cells thereby resulting in loss of the unlabeled cells.

Antigens, CD34↗
PubMedDate not supplied

Oligonucleotides in the treatment of leukemia.

The use of synthetic oligonucleotides directed towards specific genes in the therapy of leukemias has evolved rapidly over the past 5 years to early clinical trials. Undoubtedly, use of these compounds for systemic therapy and bone marrow 'purging' will escalate. Such trials will be models for the treatment of many other malignant diseases and, indeed, non-malignant diseases which may be expected to respond to eradication of a specific gene function. We are currently at the threshold of a new era of therapy which holds the promise of totally transforming clinical medicine in the next decade.

Clinical Trials as Topic↗
PubMed1990

Role of cell cholesterol in modulating antineoplastic ether lipid uptake, membrane effects and cytotoxicity.

Membrane-interactive ether lipids (EL) exert toxic and antiproliferative effects on cancer cells in vitro. They appear to be selectively more toxic to cancer cells than to normal cells and thus they are ideal candidates for bone-marrow purging procedures. However, no conclusive explanation has yet been provided for this property. We now present some data indicating that the cholesterol concentration in the incubation medium modulates EL toxicity against the HL60 leukemic cell line in vitro. Furthermore, model membranes richer in cholesterol take up EL more slowly, and cell cholesterol enrichment of HL60 cells counteracts EL biophysical membrane interaction, but not toxicity, in our experimental model. However, the K562 cell line, a leukemia line less sensitive to EL toxic action, has higher levels of cell cholesterol. Our data provide evidence to explain differences in sensitivity to EL among different cell types and contribute to the understanding of the mechanism of action of EL.

Antineoplastic Agents↗
PubMed1993

Ex vivo clonotype primer-directed gene amplification to identify malignant T cell repertoires.

A novel strategy that utilizes input genomic DNA and overcomes limitations encountered with traditional RNA reverse transcription-polymerase chain reaction (PCR) amplification methodology is described to screen for T cell receptor (TCR) repertoires. The methodology has been developed to identify individual T cell clonotypes with regard to their unique receptor beta chain variable/diversity/joining (VDJ) region gene rearrangement. The technique avoids preselection for a given antigen specificity and is therefore independent of artificial bias introduced by in vitro cell population expansion. This technique was used to detect and identify genetically of malignant clones from heterogeneous mononuclear cell populations from an array of hemato-oncological disorders, including mycosis fungoides/Sézary Syndrome, adult T cell leukemia, and large granular lymphoproliferative disease. An initial primary PCR, directed by a TCR-J beta generic primer and a complement of family-specific TCR-V beta primers, defines predominant T cell receptor variable gene usage. Use of a TCR-J beta generic primer supplants the use of a constant region primer anchor and thus eliminates the need to target mRNA. The process of variable gene screening also expedites gene sequencing. By sequencing through the VDJ juxtaposed region, i.e., the third complementarity determinant region, clonotype-specific primers are developed and used in a secondary clonotype primer-directed PCR (CPD-PCR) to detect, with extreme sensitivity and specificity, unique T cell clonal repertoires. Analysis of the products of the CPD-PCR permits the detection of a single malignant cell among one million polyclonal cells and supercedes the constraints of prior studies that provide a limited evaluation of family variable gene repertoire usage. This strategy may be applied in the detection of minimal residual disease, in surveillance after induction of disease-free states, and in analyzing the effectiveness of purging autologous bone marrow of malignant clones.

Base Sequence↗
PubMed1989

A subline of the Brown Norway myeloid leukemia in the Lewis x Brown Norway rat: in vivo growth characteristics and development of an in vitro clonogenic assay.

A subline of Brown Norway (BN) acute myelocytic leukemia (AML) which can be propagated in suspension culture (designated IPC-81) is described. Injection into Lewis x BN F1 hybrid (LBN) rats resulted in a log-linear correlation between tumor cell dose and time till death from the onset of leukemia even after multiple (greater than 16) passages in vitro. An in vitro clonogenic assay for IPC-81 colony formation (CFU-leuk) was developed with excellent cloning efficiency (55-82%). Colonies grew without the addition of specific growth factors; syngeneic spleen-conditioned medium inhibited CFU-leuk by 40%, but co-culture with untreated normal LBN rat bone marrow cells had no effect on CFU-leuk. CFU-leuk could be detected in the bone marrow 7 to 10 days before morphologic detection of leukemia in injected animals. This cell line should prove useful in the preclinical evaluation of new strategies for treating AML and evaluating new bone marrow purging methods.

Animals↗
PubMed1994

Inhibition of induced angiogenesis in a human microvascular endothelial cell line by ET-18-OCH3.

Alkyl-lysophospholipids are a group of anti-cancer compounds that have previously been shown to have the unique feature of being selectively toxic to neoplastic tissues. One of these compounds, ET-18-OCH3, has been used for purging bone marrow of cancer cells in phase I clinical trials. Tumor-induced angiogenesis has been directly correlated with tumor growth and metastasis. In this study, we examined the effect ET-18-OCH3 has on a human microvascular endothelial cell line (HMEC-1), including the following functions: angiogenesis, cell-adhesion molecule expression, and cell-junction integrity. We found that ET-18-OCH3 (in vitro) reversibly inhibited induced angiogenesis at levels that did not affect viability. At lower concentrations, ET-18-OCH3 down-regulated the expression of cell-adhesion molecules and affected the integrity of cell-to-cell junctions. This observation demonstrates this versatile family of compounds to have additional targets of action.

Antineoplastic Agents↗
PubMed1993

IgM monoclonal antibody JD118 recognizes an inducible antigen target for human-complement-mediated cytotoxicity against neoplastic B cells.

Cancer therapy using unconjugated monoclonal antibodies (mAb) has been limited by the lack of immune effector function of the mAb and antigenic modulation. JD118 is a cytotoxic murine IgM mAb with reactivity restricted to a subset of normal B cells, some monocytic series cells, and a large percentage of B cell hematopoietic neoplasms including acute and chronic leukemias and lymphomas. Specificity was determined on several hundred normal and neoplastic, hematopoietic and non-hematopoietic cells and tissues, as well as progenitor cells. JD118 was able to kill fresh human leukemias and lymphomas in the presence of human serum as a complement source with an LD50 of 100 ng/ml. At this mAb concentration, fewer than 4% of more than 10(5) available target sites were bound. Killing was not affected by changes in antigen expression observed during the cell cycle nor by loss of cell-surface targets via antigenic modulation. Cytotoxicity could be achieved with human serum diluted as low as 5%, suggesting that complement depletion in vivo should not limit activity. Autologous human serum could be used effectively as a complement source. The JD118 antigen target has not been identified, but it appears to be a glycoprotein. Up-regulation of antigen expression on normal B cells and chronic lymphocytic leukemia cells in vitro resulted in antigen-negative neoplasms becoming positive and thus targets for JD118 killing. The restricted expression, potent cytotoxic characteristics, and potential for up-regulation of its antigen make JD118 a possible candidate for ex vivo autologous bone marrow purging and in vivo therapeutic trials in patients with B cell neoplasms.

Animals↗
PubMed1993

Determination of ascorbic acid by isotachophoresis with regard to its potential in neuroblastoma therapy.

Analytical capillary isotachophoresis was used to determine ascorbic acid (AA) in different matrices (cell-free system, neuroblastoma cell extracts and urine). The system for purging bone marrow of neuroblastoma cells, including 6-hydroxydopamine (6-OHDA) and AA, was analysed with regard to the interaction of AA with 6-OHDA and its autoxidation product, hydrogen peroxide. Furthermore, analyses concerning the uptake of AA into neuroblastoma cells as well as its excretion in urine after uptake of large amounts were carried out.

Ascorbic Acid↗
PubMed1992

A novel methodology for the establishment of neuroblastoma cell lines from metastatic marrow. Expression of surface markers, neurofilaments, MDR-1 and myc proteins.

A methodology for rapid isolation of neuroblastoma (NB) cells from marrow with metastatic NB cells was developed using a cocktail of five antibodies and magnetic microspheres coated with secondary antibodies. Cells bound to microspheres were released by brief exposure to chymopapain, followed by repeated culture of released cells in serum supplemented DMEM medium and selection for adherent cells. Using this methodology, over 35 primary cell lines were obtained free of contaminating normal cells. Detailed analyses of over 14 cell lines revealed gross differences in cell phenotype, size, morphology development of neurite processes, and doubling time (40h-80 h). All cell lines expressed the 145 kDa neurofilament (NF) and a few expressed the 200 kDa NF, with very little or no expression of the 68 kDa NF. DNA analyses revealed 80% near diploid cell lines. High expression of the MDR-1 protein was detected in 6/22 of the cell lines tested.

ATP Binding Cassette Transporter, Subfamily B, Mem↗