scholarly journals Serial in vitro bone marrow fibroblast culture in human leukemia

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 589-592 ◽  
Author(s):  
T Nagao ◽  
K Yamauchi ◽  
M Komatsuda
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Colony Formation ◽  
Calf Serum ◽  
Culture Method ◽  
Fibroblast Culture ◽  
Human Leukemia ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia

Abstract Human fibroblast colony formation from bone marrow was performed in liquid culture. Fetal calf serum was used as a stimulator of the fibroblast colony formation. The colony formation took place not only in normal donors, but also in patients with acute leukemia and chronic myelocytic leukemia. At the diagnosis of the disease, significant colony suppression was observed in most cases of acute leukemia, while the number of colonies increased in half of the cases of chronic myelocytic leukemia. However, there was no correlation between the colony-forming efficiency and the initial number of peripheral platelets or bone marrow megakaryocytes that contained growth-promoting factor. The number of colonies increased after chemotherapy, recovered at the stage of complete remission, and then decreased to low levels at relapse in the patients with acute leukemia; it decreased after treatment with busulfan in the patients with chronic myelocytic leukemia. This fibroblast culture method is useful for counting fibroblast colony-forming cells in the bone marrow of human leukemia.

scholarly journals Serial in vitro bone marrow fibroblast culture in human leukemia

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 589-592 ◽  
Author(s):  
T Nagao ◽  
K Yamauchi ◽  
M Komatsuda
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Colony Formation ◽  
Calf Serum ◽  
Culture Method ◽  
Fibroblast Culture ◽  
Human Leukemia ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia

Human fibroblast colony formation from bone marrow was performed in liquid culture. Fetal calf serum was used as a stimulator of the fibroblast colony formation. The colony formation took place not only in normal donors, but also in patients with acute leukemia and chronic myelocytic leukemia. At the diagnosis of the disease, significant colony suppression was observed in most cases of acute leukemia, while the number of colonies increased in half of the cases of chronic myelocytic leukemia. However, there was no correlation between the colony-forming efficiency and the initial number of peripheral platelets or bone marrow megakaryocytes that contained growth-promoting factor. The number of colonies increased after chemotherapy, recovered at the stage of complete remission, and then decreased to low levels at relapse in the patients with acute leukemia; it decreased after treatment with busulfan in the patients with chronic myelocytic leukemia. This fibroblast culture method is useful for counting fibroblast colony-forming cells in the bone marrow of human leukemia.

scholarly journals Constitutive expression of GATA-1, EPOR, alpha-globin, and gamma-globin genes in myeloid clonogenic cells from juvenile chronic myelocytic leukemia

Blood ◽  
1995 ◽  
Vol 86 (1) ◽  
pp. 323-328 ◽  
Author(s):  
E Privitera ◽  
R Schiro ◽  
D Longoni ◽  
A Ronchi ◽  
A Rambaldi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Leukemic Cells ◽  
Globin Genes ◽  
Chronic Myelocytic Leukemia ◽  
Beta Globin ◽  
Globin Mrna ◽  
Gamma Globin ◽  
Myelocytic Leukemia ◽  
Alpha Globin

Juvenile chronic myelocytic leukemia (JCML) is a rare disorder of early childhood. Characteristic of JCML are the progressive appearance of high levels of fetal hemoglobin (HbF), reflecting a true reversion to a fetal type of erythropoiesis, and the presence of colony-forming cells able to grow in vitro spontaneously in the absence of growth factors. To better understand the relationship between the erythroid abnormalities and the leukemic process, we analyzed the expression pattern of specific genes related to erythroid differentiation--GATA-1, EPOR, alpha-globin, beta-globin, and gamma-globin genes--in JCML peripheral blood (PB) cells and in vitro-derived colonies. Northern blot analysis of PB cells from five JCML patients indicated levels of GATA-1 transcripts much higher than those usually found in other types of leukemic cells, and S1 nuclease protection assay detected significantly increased expression of gamma-globin mRNA. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of single granulocyte-macrophage colony-forming unit (CFU-GM) colonies, obtained in vitro in the absence of added growth factors from four JCML patients, detected GATA-1, EPOR, and globin (alpha and gamma) transcripts in most of the colonies tested, in contrast with control CFU-GM from normal bone marrow, which were positive only for GATA-1. Single JCML colonies were tested for the presence of two different transcripts; whereas alpha- and gamma-globin genes appeared mostly coexpressed, beta-globin mRNA was detected only in a minority of the gamma-globin-positive colonies, indicating that the leukemic pattern of hemoglobin synthesis is mainly fetal. In addition, the leukemic cells occurring during blast crisis of one of our patients displayed the typical features of a stem cell leukemia (CD34+, CD19-, CD2-, myeloperoxidase-). In this sorted CD34+ population, we detected the presence of a marker chromosome, der(12)t(3;12), previously identified in bone marrow cells at diagnosis and an expression pattern superimposable to that of the JCML colonies, consistently displaying a high gamma-globin:beta-globin mRNA ratio. The expression of erythroid markers within populations of leukemic cells, both in vivo and in vitro, supports the hypothesis that abnormal JCML erythroid cells may originate from the same mutated progenitor that sustains the growth of the leukemic cells.

scholarly journals Serial In Vitro Marrow Culture in Acute Myelocytic Leukemia

Blood ◽  
1973 ◽  
Vol 42 (5) ◽  
pp. 679-686 ◽  
Author(s):  
J. M. Bull ◽  
M. J. Duttera ◽  
E. D. Stashick ◽  
J. Northup ◽  
E. Henderson ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Clinical Remission ◽  
Colony Formation ◽  
Initial Diagnosis ◽  
Acute Myelocytic Leukemia ◽  
Normal Numbers ◽  
Myelocytic Leukemia ◽  
Large Numbers ◽  
Repeated Observation

Abstract The in vitro granulocyte colony-forming ability of bone marrow from 19 patients with acute myelocytic leukemia (AML) was studied at the time of initial diagnosis and serially. Initially 18 of the 19 patients grew scant numbers of colonies, one produced large numbers of colonies. When clinical remission was attained and leukemic blasts in the marrow were reduced to <5%, colony forming ability usually returned to normal. Eight such patients who achieved clinical remission were serially studied during remission. Normal in vitro colony formation ensued in five of the eight patients studied. Three patients did not produce normal numbers of colonies, and it was noted that these patients relapsed within 2 mo. The repeated observation of normal numbers of granulocyte colonies appears to distinguish patients whose remissions are stable from those patients who relapse quickly.

scholarly journals Erythroid Homograft Following Leukocyte Transfusion in a Patient with Acute Leukemia

Blood ◽  
1965 ◽  
Vol 26 (5) ◽  
pp. 587-596 ◽  
Author(s):  
ROBERT H. LEVIN ◽  
JACQUELINE WHANG ◽  
PAUL P. CARBONE ◽  
EMIL J. FREIREICH
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Blood Group ◽  
Acute Lymphocytic Leukemia ◽  
Lymphocytic Leukemia ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Factors Influencing ◽  
Female Donor ◽  
Treatment Of Infection

Abstract A patient with acute lymphocytic leukemia developed a functioning erythroid homograft after transfusion of leukocytes from donors with chronic myelocytic leukemia. Two donor populations were observed cytogenetically, by examination of the recipient’s marrow for sex and Philadelphia chromosomes. The cells from a female donor of different blood group eventually repopulated the entire bone marrow and were exclusively responsible for the function of the homograft. Although the donor was anemic and required periodic transfusions, her transplanted cells caused normal erythropoiesis in the recipient for a prolonged period. During this time the recipient enjoyed remission of acute leukemia but expired as a result of overwhelming infection. It is proposed that multiple transfusions of leukocytes is a useful technic for treatment of infection, for increasing knowledge of factors influencing hematopoiesis, and for studying approaches to effective immunotherapy of leukemia.

scholarly journals Improved Plasma Culture System for Production of Erythrocytic Colonies In Vitro: Quantitative Assay Method for CFU-E

Blood ◽  
1974 ◽  
Vol 44 (4) ◽  
pp. 517-534 ◽  
Author(s):  
David L. McLeod ◽  
Mona M. Shreeve ◽  
Arthur A. Axelrad
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Culture System ◽  
Fetal Calf Serum ◽  
Bone Marrow Cells ◽  
Calf Serum ◽  
Quantitative Assay ◽  
Assay Method ◽  
Wide Range

Abstract An improved plasma culture system is described for the production of erythrocytic colonies by mammalian adult hemopoietic cells in vitro under the influence of erythropoietin. The concentration of fetal calf serum in the medium used for dilution of the cells was critical for erythrocytic colony formation when low numbers of cells were plated. Optimal concentrations were found for plasma, fetal calf serum, bovine serum albumin, and L-asparagine in the culture medium, and the colony-forming efficiency was shown to depend on the concentration of erythropoietin. With erythropoietin at plateau concentration, the number of erythrocytic colonies produced was directly proportional to the number of bone marrow or spleen cells plated, over a wide range of cell concentrations. Colony numbers per culture conformed to a Poisson distribution. Thus, the improved plasma culture system may be used for the quantitative assay of CFU-E. The method is rapid (2 days), reliable, convenient, and inexpensive. Since the improved plasma culture system also supports granulocytic colony formation by bone marrow cells in the presence of conditioned medium (CSA), and the number of granulocytic colonies produced is proportional to the number of cells plated, the same hemopoietic cell suspensions can be simultaneously assayed for CFU-E and CFU-C under virtually identical conditions.

scholarly journals Mesenchymal Stem/Stromal Cells from Discarded Neonatal Sternal Tissue: In Vitro Characterization and Angiogenic Properties

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Shuyun Wang ◽  
Lakshmi Mundada ◽  
Eric Colomb ◽  
Richard G. Ohye ◽  
Ming-Sing Si
Keyword(s):  
Bone Marrow ◽  
Cardiac Surgery ◽  
Stromal Cells ◽  
Umbilical Vein ◽  
Culture Method ◽  
Tissue Perfusion ◽  
Explant Culture ◽  
Critical Congenital Heart Disease ◽  
Surface Marker Expression

Autologous and nonautologous bone marrow mesenchymal stem/stromal cells (MSCs) are being evaluated as proangiogenic agents for ischemic and vascular disease in adults but not in children. A significant number of newborns and infants with critical congenital heart disease who undergo cardiac surgery already have or are at risk of developing conditions related to inadequate tissue perfusion. During neonatal cardiac surgery, a small amount of sternal tissue is usually discarded. Here we demonstrate that MSCs can be isolated from human neonatal sternal tissue using a nonenzymatic explant culture method. Neonatal sternal bone MSCs (sbMSCs) were clonogenic, had a surface marker expression profile that was characteristic of bone marrow MSCs, were multipotent, and expressed pluripotency-related genes at low levels. Neonatal sbMSCs also demonstrated in vitro proangiogenic properties. Sternal bone MSCs cooperated with human umbilical vein endothelial cells (HUVECs) to form 3D networks and tubes in vitro. Conditioned media from sbMSCs cultured in hypoxia also promoted HUVEC survival and migration. Given the neonatal source, ease of isolation, and proangiogenic properties, sbMSCs may have relevance to therapeutic applications.

Preclinical Evaluation of a Bone-Marrow Autograft Culture Procedure for Generating Lymphokine-Activated Killer Cells in Vitro

1992 ◽  
Vol 3 (suppl b) ◽  
pp. 123-127 ◽  
Author(s):  
Hans-Georg Klingemann ◽  
Heather Deal ◽  
Dianne Reid ◽  
Connie J Eaves
Keyword(s):  
Bone Marrow ◽  
Calf Serum ◽  
Cell Activity ◽  
Hematopoietic Cells ◽  
Lak Cells ◽  
High Dose ◽  
Lymphokine Activated Killer Cells ◽  
Lak Cell

Despite the use of high dose chemoradiotherapy for the treatment of acute leukemia. relapse continues to be a major cause of death in patients given an autologous bone marrow transplant. Further augmentation of pretransplant chemotherapy causes life threatening toxicity to nonhematopoietic tissues and the effectiveness of currently available ex vivo purging methods in reducing the relapse rate is unclear. Recently, data from experimental models have suggested that bone marrow-derived lymphokine (IL-2)-activated killer (BM-LAK) cells might be used to eliminate residual leukemic cells both in vivo and in vitro. To evaluate this possibility clinically, a procedure was developed for culturing whole marrow harvests with IL-2 prior to use as autografts, and a number of variables examined that might affect either the generation of BM-LAK cells or the recovery of the primitive hematopoietic cells. The use of Dexter long term culture (LTC) conditions, which expose the cells to horse serum and hydrocortisone. supported LAK cell generation as effectively as fetal calf serum (FCS) -containing medium in seven-day cultures. Maintenance of BM-LAK cell activity after a further seven days of culture in the presence of IL-2 was also tested. As in the clinical setting. patients would receive IL-2 in vivo for an additional week immediately following infusion of the cultured marrow autograft. Generation ofBM-LAK activity was dependent on the presence of IL-2 and could be sustained by further incubation in medium containing IL-2. Primitive hematopoietic cells were quantitated by measuring the number of in vitro colony-forming progenitors produced after five weeks in secondary Dexter-type LTC. Maintenance of these 'LTC-initiating cells' was unaffected by lL-2 in the culture medium. These results suggest that LAK cells can be generated efficien tly in seven-day marrow autograft cultures containing IL-2 under conditions that allow the most primitive human hematopoietic cells currently detectable to be maintained.

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