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PubMed1990

Development of T cells in vitro from precursors in mouse bone marrow.

Bone marrow cells from 6- to 8-week-old athymic nude mice were depleted of nylon-wool adherent cells and cultured in vitro at low cell numbers (300 cells/well) in medium supplemented with a supernatant from a thymoma cell line. About 1% of cultured cells grew. Pooled cultures contained cells expressing CD3 (52%), CD4 (37%), CD8 (11%), Thy 1.2 (72%), MAC-1 (43%) and J11d (86%) but no cells expressing sIg. They also contained cells expressing mRNA for the alpha, beta, gamma, and delta chains of the T cell receptor as assessed with C region probes using a sensitive dot blot assay. These cells appear to develop from progenitors which are CD3-. When pooled Day 10 cultures were depleted of nylon-wool adherent cells, the remaining cells were nearly all J11d+, Thy 1.2+, MAC-1-, CD3+, and either CD4+CD8+; CD4+CD8-; CD4-CD8+, or CD4-CD8-; i.e., their surface marker patterns were reminiscent of those of thymocytes. We conclude that our culture system is enabling bone marrow precursors to commence differentiation down the T cell lineage in the absence of a thymic environment.

Animals↗

PubMed1997

Bone marrow imaging of bone marrow transplanted multiple myeloma patients.

Eighteen bone marrow transplanted multiple myeloma patients had imaging studies on 24 occasions with radiography as well as bone and bone marrow scintigraphy within 2 months. Twelve of the radionuclide bone marrow studies were performed with Tc-99m human serum albumin colloid and 12 were performed with a Tc-99m tagged monoclonal antigranulocyte antibody. The total detection rate of bone marrow lesions increased by 5% when the findings on bone marrow scintigraphy were combined with the findings and at radiography bone scintigraphy. For lesions in the spine and sacrum, the increase was 25% and 33% respectively, including patients with focal radiotherapy. Peripheral red bone marrow expansion was noted in 17 patients. In a comparison of Mab and Tc-99m HSA colloid imaging, Mab resulted in a higher bone marrow to soft tissue uptake and to a much smaller part of the skeleton being obscured by liver and spleen uptake. It is concluded that bone marrow imaging is valuable for showing red bone marrow distribution. It thereby shows possible sites for malignant lesions; it also shows that Mab imaging is superior to Tc-99m HSA colloid imaging in bone marrow transplanted multiple myeloma patients.

Adult↗

PubMed1995

Use of cryopreserved bone marrow in allogeneic bone marrow transplantation.

Use of cryopreserved donor bone marrow may facilitate scheduling of allogeneic bone marrow transplantation (BMT) by affording independence of a fixed time for bone marrow donation. The potential risk of damage to hematopoietic stem cells by cryopreservation resulting in delayed engraftment or graft failure has to be taken into account, however. To address these issues, the outcome of 19 matched related BMT (1992-94) performed with cryopreserved donor bone marrow was analyzed and compared with 19 related BMT (1990-93) receiving fresh donor bone marrow (control group). Time to engraftment of patients receiving cryopreserved bone marrow was not different from the control group (ANC > 0.2 x 10(9)/l 15.5 +/- 3.8 days vs 15.8 +/- 5.0 days; ANC > 0.5 x 10(9)/l 17.3 +/- 4.1 days vs 17.9 +/- 5.0 days, respectively). We did not find the previously described trend toward a lower incidence of acute GVHD in patients receiving cryopreserved bone marrow compared with patients receiving fresh bone marrow (acute GVHD > or = II 78 vs 64%). Furthermore, the two groups did not differ in the incidence of chronic GVHD (55 vs 38%) or day 100 survival (74 vs 68%). We conclude that in allogeneic BMT cryopreserved bone marrow cells can be safely used without jeopardising or prolonging time to engraftment.

Adolescent↗

PubMed1994

Human herpesvirus 6 (HHV-6) isolation from bone marrow: HHV-6-associated bone marrow suppression in bone marrow transplant patients.

These studies tested the hypothesis that human herpesvirus 6 (HHV-6) can cause posttransplant bone marrow (BM) suppression in BM transplant (BMT) patients. Fifteen adult patients who received T-lymphocyte-depleted, allogeneic BM transplants and who developed posttransplant BM suppression were studied. Detailed chart reviews were used to divide the patients into two groups: (1) those with diagnosed BM suppression (DBMS) and (2) those with idiopathic BM suppression (IBMS). BM aspirates obtained from the patients at the onset of BM suppression were subjected to an HHV-6 isolation procedure using mitogen-stimulated blood mononuclear cells. BM specimens obtained from another population of BMT patients solely to document engraftment irrespective of their BM function were also subjected to the HHV-6 isolation procedure as controls. HHV-6 was isolated from 6 of 15 BM samples from BMT patients with BM suppression. BM samples from patients with IBMS were more likely to be positive for HHV-6 than those from patients with DBMS (P < .01). Also, HHV-6-positive BM were significantly more likely (P < .05) to come from patients with suppression of more than one BM lineage than HHV-6-negative BM. Finally, samples of BM from an unselected series of BMT patients studied without regard to their BM function were less likely (P < .01) to be positive for HHV-6 than patients with IBMS.

Adult↗

PubMedDate not supplied

Initial experience with dynamic MR imaging in evaluation of normal bone marrow versus malignant bone marrow infiltrations in humans.

The purpose of this study was (a) evaluation of dynamic contrast-enhanced MR imaging of normal bone marrow versus malignant bone marrow infiltrations in patients with proven B-cell-type chronic lymphocytic leukemia (B-CLL) and (b) correlation with the clinical stage according to Binet (stages A, B, C) and response to therapy. Bone marrow imaging of the lumbar spine, pelvis, and proximal femurs was performed at 1.5 T in 45 patients without known malignancy and in 30 patients with B-CLL. The differences between opposed-phase and in-phase dynamic gradient-echo sequences before and up to 10 minutes after intravenous application of .1 mmol/kg body weight of gadolinium-diethylenetriamine penta-acetic acid (Gd-DTPA) were evaluated in normal bone marrow. The contrast-enhancement patterns of normal and malignant bone marrow were compared using the opposed-phase dynamic gradient-echo sequence. Ten of the patients with bone marrow infiltrations (Binet stage C) additionally underwent MR imaging follow-up during therapy. Opposed-phase gradient echo sequences demonstrated a signal decrease of normal bone marrow, and in-phase gradient echo sequences demonstrated a signal increase of normal bone marrow after administration of Gd-DTPA. The dynamic signal intensity time courses differed significantly (P < .05) between Binet stages B and C and controls as well as among the three Binet stages of B-CLL. In the 10 patients followed during therapy, MR imaging sensitively demonstrated response (n = 6), nonresponse (n = 2), or relapse after initial response (n = 2). In out-of-phase imaging, both normal bone marrow and initial bone marrow infiltration in CLL stage Binet A show signal decrease after administration of contrast agent, whereas there is increase in signal intensity in higher-grade bone marrow infiltration in Binet stage B or C disease. The signal loss of normal bone marrow in out-of-phase imaging is a phase effect rather than a T2* effect. The differentiation of initial from higher-grade bone marrow infiltration on out-of-phase images relies solely on a shift in the fat/water ratio.

Adult↗

PubMed1994

B-cell precursor bone marrow reconstitution after bone marrow transplantation.

Bone marrow transplantation is characterized by a prolonged period of humoral immunodeficiency in which many patients have abnormal circulating B-cell subsets, and oligoclonal and monoclonal gammapathies. In this study we examine B-cell precursor reconstitution in the post-transplantation marrow. Within 1 month after transplantation there is a marked increase in the percentage of immature B cells (to 80% of marrow lymphocytes), which can persist for more than 1 year. The increase in B-cell precursors is seen in both adults and children and appears to be independent of age. These cells have a normal precursor B-cell surface antigenic phenotype (CD19+, CD10+, CD20 negative to dim) and generally express very little CD34. No monoclonal or oligoclonal immunoglobulin gene rearrangements are detected in these cells, which enables them to be easily distinguishable from common precursor B-cell acute lymphocytic leukemia lymphoblasts.

Adolescent↗

PubMed2006

Development of the osteoblast phenotype of serial cell subcultures from human bone marrow.

Bone marrow cells have been used for testing biocompatibility of bone substitute materials that would be applied in maxillofacial and orthopedic surgeries. However, it remains unclear whether cells in serial subcultures retain the ability to differentiate into osteoblasts. The purpose of this study was to compare the development of osteoblast phenotype of serially passaged cells from human bone marrow. Cells from first to third passage were cultured (2 x 10(4) cells/well) in supplemented culture medium. Cells were incubated at 37 degrees C in a humidified atmosphere of 5% CO2 and 95% air. Cell attachment was assessed at 4 and 24 h. At 7, 14 and 21 days, cell proliferation, cell viability, total protein content and alkaline phosphatase (ALP) activity were evaluated. Bone-like formation was evaluated at 14 and 21 days. Data were compared by two-way ANOVA and Duncan's multiple range test. Cell attachment, cell viability and total protein content were not affected by serial subcultures. However, serial subcultures did interfered negatively with osteoblast differentiation as shown by osteoblast parameters observed in second and third subcultures, such as continuous cell proliferation, lower ALP activity and bone-like formation in comparison to first subculture. Therefore, it is important to evaluate cell ability to growth and differentiate before selecting the cell population for studies that investigate the biocompatibility of materials to replace bone tissue.

Adolescent↗

PubMed2005

Comparison of relative value of bone marrow aspirates and bone marrow trephine biopsies in the diagnosis of solid tumor metastasis and Hodgkin lymphoma: institutional experience and literature review.

CONTEXT: Bone marrow aspirates as well as bone marrow trephine biopsies are frequently performed to assess whether there is marrow involvement by a malignancy. Numerous reports differ in the relative value of these 2 procedures and fail to provide concise guidelines that can help choose the appropriate technique in this clinical situation. OBJECTIVE: To compare the relative value of aspirates and trephine biopsies in the diagnosis of solid tumor metastasis and Hodgkin lymphoma. In addition, we correlate our findings with those of the literature to provide a concise practice guideline. DESIGN: Sixty-six cases showing bone marrow involvement by solid tumor and Hodgkin lymphoma in bone marrow aspirates, bone marrow trephine biopsies, or both were included in the study. The diagnosis and findings made on aspirates were compared with those made on trephine biopsies in each case. RESULTS: In those cases where both aspirate and trephine biopsy were available for evaluation, there was a 22% positive correlation in the findings on aspirates and trephine biopsies. The correlation between aspirates and trephine biopsies was highest in cases of small cell carcinoma of the lung (3/11, or 36.3%) followed by breast carcinoma (7/20, or 35%), prostate carcinoma (1/9, or 11.1%), and Hodgkin lymphoma (1/20, or 5%). Two of 5 cases from the miscellaneous category demonstrated simultaneous involvement of aspirate and trephine biopsy by a gastric carcinoma as well as an adrenal gland carcinoma. CONCLUSIONS: Bone marrow aspirate and bone marrow trephine biopsy should both be performed in patients with proven or suspected malignancies where staging may affect management. However, bone marrow aspirate has only a minimal role, if any, in detecting bone marrow involvement by Hodgkin lymphoma. In cases of breast carcinoma, small cell carcinoma of lung, and prostate carcinoma, aspirate evaluation may confirm trephine biopsy results or, more rarely, provide the sole confirmation of the malignancy.

Biopsy↗

PubMed1987

Alcohol-induced bone marrow damage. A bone marrow study in alcohol-dependent individuals.

Bone marrow biopsies from 30 alcohol-dependent individuals hospitalized for detoxification were investigated. Typical alcohol-induced bone marrow changes were found and served to define alcohol-induced bone marrow damage as a nosological entity. The findings took the form of heightened ineffective erythropoiesis associated with impaired iron utilization, vacuolated proerythroblasts, multinuclear erythroblasts, megaloblasts and iron-containing plasma cells as well as vacuolated precursor cells of the granulocytopoietic series. In the differential diagnosis, alcohol-induced bone marrow damage is to be distinguished from the myelodysplastic syndrome of the RA and RARS form. Alcohol-induced bone marrow damage is reversible. Bone marrow cell cultures performed in our cases are normal, showing that the toxic defect probably does not reside in the stem cell but is more peripheral. Normal bone marrow cell culture may be a typical feature of alcohol-induced bone marrow damage.

Adolescent↗

PubMed1996

Histopathology of bone marrow after allogeneic bone marrow transplantation for chronic myeloid leukaemia.

The histopathology of bone marrow after allogeneic bone marrow transplantation for chronic myeloid leukaemia (CML) was studied in 20 bone marrow biopsy specimens from 14 patients. Biopsies were performed at day 30 post-transplantion (5 biopsies), between day 90 and day 100 (14 biopsies), and one year post-transplantation (1 biopsy). Eight biopsies taken at day 30 post-transplantation for lymphoproliferative disorder were studied as controls. Granulopoiesis was predominant in all day 30 bone marrows in patients treated for CML. In 2 cases, rejection was suspected because of marrow hypoplasia at day 30: one case developed acute leukaemia soon after persistant hypoplasia at day 90 while the evolution of the other case was good. Two day 30 biopsies showed granulocytic hyperplasia but follow-up proved complete remission of CML. Thus, in our series, early biopsies at day 30 did not have any prognostic value. Twelve cases at day 90-100 showed little modifications, such as minor dyserythropoiesis and dysmegakaryopoiesis. One case showed granulocytic hyperplasia at day 100 but bone marrow was normal one year later and the patient in complete remission. The haematopoietic reconstitution after allogeneic bone marrow transplantation may present a transient granulocytic hyperplasia that must not be diagnosed as a persistence of CML.

Adult↗

PubMed1993

The effect of sequence and time interval between cyclophosphamide and total body irradiation on lung and bone marrow damage following bone marrow transplantation in mice.

The compromise between bone marrow killing effect and toxicity (mainly on the lungs) of the conditioning protocol used prior to bone marrow transplantation (BMT) determine to a great extent the final outcome. In search of an optimal balance between minimum lung damage and maximum bone marrow cell kill, we have tested the effect of varying the sequence and time interval between cyclophosphamide (CTX) and total body irradiation (TBI). CTX was administered almost concomitantly with TBI (i.e., 15 min before TBI) or 1-7 days before or after TBI. Lung damage was assessed by the lethality (LD) of the mice between day 28 and day 180 after treatment, bone marrow damage by the LD of the mice between day 7 and day 28 after treatment and by the spleen colony assay. Mice chosen for lung damage testing were rescued from death due to bone marrow ablation by transplantation with syngeneic marrow cells. CTX potentiated radiation damage in both bone marrow and lungs. The effect on the bone marrow was greater when CTX was given after TBI than when given before TBI and this effect was significantly more than additive when the interval between the two agents was 3 days. Lung toxicity, on the other hand, was greater when CTX was given before TBI than when given after TBI. A therapeutic gain factor (TGF) was estimated by dividing the dose enhancement ratio (DEF) of bone marrow damage over the DEF of lung damage at all the time intervals studied. The results were consistently higher when CTX was given after TBI than when given before.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1996

Purging of bone marrow in autologous bone marrow transplantation for non-Hodgkin's lymphoma: a case-matched comparison with unpurged cases by the European Blood and Marrow Transplant Lymphoma Registry.

PURPOSE: The use of in vitro purging of bone marrow in autologous bone marrow transplantation (ABMT) for non-Hodgkin's lymphoma (NHL) has been a controversial issue; its benefit is as yet unproven. Its effect on the clinical outcome of ABMT in these patients is still unclear. We look at this issue using data from the European Blood and Marrow Transplant (EBMT) Lymphoma Registry. PATIENTS AND METHODS: Seventeen hundred twenty-six patients with NHL have been reported to the EBMT registry, of whom 270 had bone marrow purged at transplant. Two hundred twenty-four of these patients were compared with a case-matched group of 224 unpurged patients who had undergone the same procedure. The case matching was made following selection of the main prognostic factors for progression-free survival (PFS) by multivariate analysis. Response, complications, and outcome in ABMT were analyzed. RESULTS: Time to hematologic engraftment, response to ABMT, and number of procedure-related deaths were similar in purged and unpurged patients. The overall survival (OS) rate was 54% at 5 years in purged patients and 48.3% in unpurged patients (P = .1813). The PFS rate was 44.3% and 44.6%, respectively (P = .1961). Patterns of relapse, including bone marrow relapse, were similar in both groups. Patients with low-grade lymphoma did not have a significantly improved PFS if the bone marrow was purged (P = .1757); however, they did have a significantly improved OS (P = .00184). This increased OS was found to be associated with non-totalbody irradiation (TBI) conditioning and also with the purged patients undergoing transplantation at large transplant centers (P = .0016). CONCLUSION: Purging of bone marrow in ABMT for NHL does not affect the rate of hematologic engraftment or risk of procedure-related death (PRD). There is no significant difference in PFS for patients whose bone marrow is purged as compared with unpurged.

Adolescent↗

PubMed1985

Erythrocyte depletion of ABO-incompatible bone marrow.

Bone marrow transplantation results in the infusion of 150 to 360 ml of erythrocytes. While this poses no problem to a recipient of marrow from an ABO-compatible donor, it clearly represents a serious risk of intravascular hemolysis for the recipient of an ABO-incompatible marrow. Thus, there is a need for removing incompatible erythrocytes from the marrow preparation. We removed erythrocytes from the marrow preparations by automated centrifugation. The erythrocyte-depleted marrow (EDM) contained a mean of 5 ml erythrocytes, representing an average reduction of 98%. The mean mononuclear cell recovery was 88%, resulting in a mean infusion of 0.6 X 10(8) cells/Kg of recipient's body weight in a final average volume of 155 ml. EDM was infused into 22 ABO-incompatible marrow recipients (21 patients with hematologic malignancies and one patient with aplastic anemia) without clinical evidence of hemolysis. The isohemagglutinin titers of recipients ranged from 4 to 4096 and were not lowered prior to infusion. Engraftment (i.e., recovery of peripheral leukocyte and platelet counts) and incidence of graft versus host disease were similar to those observed in recipients of ABO-compatible marrow transplantation. Erythrocyte engraftment was significantly delayed in only one patient who had a high isohemagglutinin titer. The post-transplantation red cell requirement was increased in EDM recipients: 9 units compared to 6 units in ABO-compatible bone marrow transplanted patients with neither hemolysis nor interference with successful engraftment.

ABO Blood-Group System↗

PubMed1988

[In vitro manipulation of bone marrow cells for bone marrow transplantation].

Recently, in vitro manipulations of bone marrow cells have been developed for the prevention of relapse in autologous bone marrow transplantation (BMT) and for the prevention of graft versus host disease in allogeneic BMT. Two methods were mainly employed clinically for the depletion of cells from transplanted bone marrow cells. Firstly, bone marrow cells were treated in vitro with monoclonal antibodies reactive to leukemia cells or T cells using complement, immunotoxin or magnetic bead. Secondly, bone marrow cells were incubated with cancer drugs such as 4-HC or mafosfamide. Our results and other reports using in vitro purging of leukemia or lymphoma cells suggest that these autologous BMTs are effective modes of cancer therapy in patients with hematological malignancies.

Acute Disease↗

PubMed1991

Stromal cell growth and blast colony-forming cells in AML bone marrow.

Bone marrow cells (BMC) from normal donors and from patients with acute myeloid leukaemia (AML) were cultured. Growth kinetics and the efficiency of stromal layers in supporting the adhesion of normal blast-colony forming cells (BL-CFCs) were studied. BMC from treated AML patients formed confluent stromal layers faster than normal BMCs. BL-CFC binding capacity of normal and AML stromal layers did not differ: on normal stromal layers 67.3-147, on AML stromal layers 63-117 colonies per 5 x 10(5) plastic non-adherent BMC were formed. The amount and/or binding capacity of BL-CFCs was found to be normal in two AML patients in complete remission, while a significantly reduced number and/or binding capacity of BL-CFCs was found in AML non-treated patients and in patients within 4 weeks after the last cytostatic course.

Blast Crisis↗

PubMed1994

Comparison of the phenotype and clonogenicity of normal CD34+ cells from umbilical cord blood, granulocyte colony-stimulating factor-mobilized peripheral blood, and adult human bone marrow.

Bone marrow (BM) is most frequently used to transplant hematopoietic progenitor cells, but umbilical cord blood (UCB) and mobilized peripheral blood (PB) provide alternative sources of progenitor cells for transplantation. To study whether the clonogenicity and phenotype of progenitor cells vary between the compartments, CD34+ cells from UCB, mobilized PB, and BM were analyzed for in vitro colony formation and characterized by immunophenotyping for several lineage-associated and maturation-related cell surface molecules. We found that circulating CD34+ cells, either from PB after granulocyte colony-stimulating factor (G-CSF) mobilization or from UCB, contained a large proportion of cells (86-96%) with myeloid cell-associated molecules (CD33 and CD13) and clonogenic cells (colony-forming unit-granulocyte-macrophage and burst-forming unit-erythrocyte) in excess of BM CD34+ cells. Further, UCB and PB CD34+ cells contained > or = 3% cells with a phenotype associated with immature (HLA-DR- and CD38-) progenitor cells, which was comparable to what is found among BM CD34+ cells. The proportion of CD34+ cells in UCB expressing the B cell-associated molecules (CD10 and CD19) was comparable to that found in mobilized PB (< or = 5%) but significantly lower than for BM CD34+ cells (19-24%). Further, we observed a higher proportion of CD34+ cells expressing the T cell-associated molecule CD7+ in UCB (7%) compared with both PB and BM (3-4%). In general, circulating CD34+ cells, either from UCB or G-CSF-mobilized PB, display largely the same phenotypic profile and clonogenicity, being different from resident CD34+ cells in BM.

ADP-ribosyl Cyclase↗

PubMed1981

[Cytogenetics in bone marrow transplantations. I. Bone marrow aplasia].

A bone marrow transplantation has been carried out in a patient with bone marrow aplasia. Besides the cytochemical and haematological tests, the cytogenetic analysis has been performed to check whether the transplantation was successful. In this case the donor was the patient's sister, so that the presence of the chimere has been used as a criteria for judging the conditions of the transplantation. the cytogenetic analysis, furthermore, is suitable to detect other chromosome abnormalities, which can represent a condition of instability of the transplanted cells and are probably an early expression of the transplanted bone marrow.

Adolescent↗

PubMed2002

Bone marrow and bone: a functional unit.

Bone and bone marrow, although often regarded as separate systems, function as a single unit. Cells in the bone marrow are the precursors of bone remodelling cells and exert an important regulatory role both on their own development and the remodelling process, acting as mediators for the effects of systemic and local factors. Other cells, such as immune cells and megakaryocytes, also contribute to the regulation of bone cell development and activity. Many diseases that affect the bone marrow have profound effects on bone, involving interactions between abnormal and normal marrow cells and those of bone. Although recent advances in bone physiology have produced new insights into the relationship between bone marrow and bone cells, much remains to be learnt about the mechanisms by which marrow and bone act in synergy to regulate bone remodelling, both in health and disease.

Aging↗