The wall of the chick embryo aorta harbours M-CFC, G-CFC, GM-CFC and BFU-E

Development ◽  
1988 ◽  
Vol 102 (2) ◽  
pp. 279-285 ◽  
Author(s):  
F. Cormier ◽  
F. Dieterlen-Lievre
Keyword(s):  
Bone Marrow ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Avian Embryo ◽  
Erythroid Progenitors ◽  
Haemopoietic Stem Cells ◽  
Different Types ◽  
Haemopoietic Cells ◽  
Very High

In the 3- to 4-day avian embryo, after the first wave of haemopoiesis which derives in the yolk sac from haemopoietic stem cells formed in situ, haemopoietic cells emerge in an intraembryonic site, the wall of the aorta. In this paper, we demonstrate that this site harbours M-CFC, G-CFC, GM-CFC and late and early BFU-E. In serum-free medium, the growth of M-CFC and GM-CFC was strictly dependent on CSF present in fibroblast-conditioned medium (FCM). The growth of G-CFC was improved when FCM was replaced by a minute quantity of chicken and fetal calf serum. Like erythroid progenitors from bone marrow, BFU-E detected here required anaemic chicken serum to differentiate into haemoglobinized cells. The frequency of the different types of haemopoietic progenitors in the aortic population was very high: 80 M-CFC, 25 G-CFC, 4 GM-CFC and 70 BFU-E for 12,500 aorta cells, i.e. two to eight times more frequent than in the bone marrow population, depending on the type of progenitors.

scholarly journals Fetal calf serum contains activities that induce fetal hemoglobin in adult erythroid cell cultures

Blood ◽  
1990 ◽  
Vol 75 (9) ◽  
pp. 1862-1869 ◽  
Author(s):  
P Constantoulakis ◽  
B Nakamoto ◽  
T Papayannopoulou ◽  
G Stamatoyannopoulos
Keyword(s):  
Cell Cultures ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Fetal Hemoglobin ◽  
Erythroid Cell ◽  
Erythroid Progenitors ◽  
Globin Mrna ◽  
Erythroid Progenitor ◽  
Fetal Sheep ◽  
Gamma Globin

Abstract Cultures of peripheral blood or bone marrow erythroid progenitors display stimulated production of fetal hemoglobin. We investigated whether this stimulation is due to factors contained in the sera of the culture medium. Comparisons of gamma/gamma + beta biosynthetic ratios in erythroid colonies grown in fetal calf serum (FCS) or in charcoal treated FCS (C-FCS) showed that FCS-grown cells had significantly higher gamma/gamma + beta ratios. This increase in globin chain biosynthesis was reflected by an increase in relative amounts of steady- state gamma-globin mRNA. In contrast to its effect on adult cells, FCS failed to influence gamma-chain synthesis in fetal burst forming units- erythroid (BFU-E) colonies. There was a high correlation of gamma- globin expression in paired cultures done with C-FCS or fetal sheep serum. Dose-response experiments showed that the induction of gamma- globin expression is dependent on the concentration of FCS. These results indicate that FCS contains an activity that induces gamma- globin expression in adult erythroid progenitor cell cultures.

On the origin of haemopoietic stem cells in the avian embryo: an experimental approach

Development ◽  
1975 ◽  
Vol 33 (3) ◽  
pp. 607-619
Author(s):  
par Françoise Dieterlen-Lievre
Keyword(s):  
Stem Cells ◽  
Blood Cell ◽  
Experimental Approach ◽  
Developmental Stages ◽  
Yolk Sac ◽  
Avian Embryo ◽  
Cell Nuclei ◽  
Haemopoietic Stem Cells ◽  
Cell Series ◽  
Haemopoietic Cells

It is currently accepted that stem cells of the definitive blood cell lines originate from the yolk-sac blood islands. Experiments were devised to examine the validity of this theory in the avian embryo. These involved grafting two-day-old quail embryos on to chick yolk-sacs of comparable developmental stages, i.e. before or shortly after the establishment of vascularization.The conclusions of the experiments are based on the possibility of distinguishing chick cell nuclei from those of the quail. In the developing haemopoietic organs (spleen and thymus) of quail embryos grafted on to the chick and subsequently incubated for 6–11 days, all cells, whether belonging to the granulopoietic, erythropoietic or lymphopoietic series, are of quail type. Thus these organs have not been colonized by chick stem cells. On the other hand, coelomic graft experiments show that the development of these organs is indeed dependent on an extrinsic colonization by haemopoietic cells; quail spleen or thymus rudiment, developing in the coelom of a chick, is populated by chick cells. Thus no incompatibility which would prevent heterospecific colonization exists in this system. It is concluded that haemopoietic stem cells of the definitive blood cell series originate from some source other than the yolk-sac, and that this source must be intra-embryonic.

Modification of Gene Expression in Mesenchymal Stromal Cells of the Leukemia Patients during Chemotherapy

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5065-5065
Author(s):  
Tamara Sorokina ◽  
Irina Shipounova ◽  
Alexey Bigildeev ◽  
Nina I. Drize ◽  
Larisa A. Kuzmina ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Acute Leukemia ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Leukemic Cell ◽  
Expression Level ◽  
Leukemic Cells ◽  
Disease Manifestation ◽  
Relative Expression Level

Abstract Background In patients with acute leukemia the stromal microenvironment is deeply modified. Disturbances in signaling pathways, genetic abnormalities and functional changes in mesenchymal cells of these patients have been previously described. Chemotherapy also affect stromal progenitor cells. A damaged microenvironment might impair hematopoiesis in acute leukemia patients. Aims To investigate the relative expression level in MMSCs and CFU-Fs, derived from the bone marrow (BM) of acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) patients before and over the course of chemotherapy. Methods 54 newly diagnosed cases (33 AML, 21 ALL) were involved in the study after informed consent. BM was aspirated prior to any treatment (time-point 0) and at days 37, 100 and 180 since the beginning of treatment of acute leukemia. MMSCs were cultured in aMEM with 10% fetal calf serum, CFU-Fs, in aMEM with 20% fetal calf serum. The relative expression level (REL) of different genes was measured by TaqMan RQ-PCR. As a control MMSCs and CFU-Fs from 88 healthy donors were used. Results At the time of the disease manifestation the analysis of gene expression in MMSCs from acute leukemia patients revealed a significant increase in the REL of genes which regulate immune system responses and thereby can influence on the leukemic cell proliferation and migration (IL-6, IL-8, IL-1b and IL-1R1) (Pic.1). Also at the time of the diagnosis an increase in REL of genes, that are responsible for hematopoiesis regulation, was observed. For example, the REL of CSF1 that can influence on leukemic cells proliferation was increased at the disease manifestation and became normal during the treatment. The same dynamics was observed in the REL of JAG1 that has an antiapoptotic effect on leukemic cells. The REL of LIF had been also significantly increased at the disease manifestation, reflecting the efforts of MMSCs to inhibit leukemic proliferation. Chemotherapy affected REL of the studied genes differently. The treatment lead to the downregulation of IGF, TGFB1 and TGFB2 (Pic.2). As far asTGFB1 and 2 inhibit the differentiation of mesenchymal stem cells, and IGF is associated with myelodysplastic changes in elderly bone marrow, so their downregulation may refer to the effectiveness of therapy. The REL of genes regulating MMSC proliferation (PDGFRa and PDGFRb, FGF2, FGFR1 and 2) increased during chemotherapy. Exploring cell adhesion molecules, the decrease in the REL of their encoding genes was observed. As far as VCAM facilitate the leukemic cell extravasation and ICAM was shown to depress the Th17 cell differentiation, the down-regulation of their genes may reflect the microenvironment restoration. The influence of chemotherapy lead to decrease in REL of genes, associated with MMSCs differentiation (BGLAP and SOX9 (Pic.3)), reflecting the mechanism of the blocking of MMSCs migration and differentiation under the stress conditions. The alterations of bone marrow stroma were more pronounced in patients who didn't achieve remission. The REL of 9 genes was studied in CFU-F colonies. There were no differences in gene expression in CFU-Fs before the treatment, except for an increase in the REL of PPARg in acute leukemia CFU-Fs. During the treatment, a decrease in the REL of SPP1 and an increase in the REL of FGFR1 and 2 were observed. Conclusion Therefore, chemotherapy used does not impair the functional ability of MMSCs and CFU-Fs, but influence on their gene expression profile. The two types of precursors are affected differently, indicating their different differentiation level and functions. Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3. Disclosures No relevant conflicts of interest to declare.

scholarly journals Reduction of erythroid progenitors in protein–energy malnutrition

2007 ◽  
Vol 97 (2) ◽  
pp. 307-314 ◽  
Author(s):  
Primavera Borelli ◽  
Solange Blatt ◽  
Juliana Pereira ◽  
Beatriz Beutler de Maurino ◽  
Maristela Tsujita ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Binding Capacity ◽  
Control Diet ◽  
Protein Energy Malnutrition ◽  
Erythroid Progenitors ◽  
Protein Energy ◽  
Colony Forming Units ◽  
Macrophage Colony ◽  
Hb Concentration ◽  
Haemopoietic Cells

Protein–energy malnutrition is a syndrome in which anaemia together with multivitamin and mineral deficiency may be present. The pathophysiological mechanisms involved have not, however, yet been completely elucidated. The aim of the present study was to evaluate the pathophysiological processes that occur in this anaemia in animals that were submitted to protein–energy malnutrition, in particular with respect to Fe concentration and the proliferative activity of haemopoietic cells. For this, histological, histochemical, cell culture and immunophenotyping techniques were used. Two-month-old male Swiss mice were submitted to protein–energy malnutrition with a low-protein diet (20 g/kg) compared with control diet (400 g/kg). When the experimental group had attained a 20 % loss of their original body weight, the animals from both groups received, intravenously, 20 IU erythropoietin every other day for 14 d. Malnourished animals showed a decrease in red blood cells, Hb concentration and reticulocytopenia, as well as severe bone marrow and splenic atrophy. The results for serum Fe, total Fe-binding capacity, transferrin and erythropoietin in malnourished animals were no different from those of the control animals. Fe reserves in the spleen, liver and bone marrow were found to be greater in the malnourished animals. The mixed colony-forming unit assays revealed a smaller production of granulocyte–macrophage colony-forming units, erythroid burst-forming units, erythroid colony-forming units and CD45, CD117, CD119 and CD71 expression in the bone marrow and spleen cells of malnourished animals. These findings suggest that, in this protein–energy malnutrition model, anaemia is not caused by Fe deficiency or erythropoietin deficiency, but is a result of ineffective erythropoiesis.

scholarly journals Fetal calf serum contains activities that induce fetal hemoglobin in adult erythroid cell cultures

Blood ◽  
1990 ◽  
Vol 75 (9) ◽  
pp. 1862-1869
Author(s):  
P Constantoulakis ◽  
B Nakamoto ◽  
T Papayannopoulou ◽  
G Stamatoyannopoulos
Keyword(s):  
Cell Cultures ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Fetal Hemoglobin ◽  
Erythroid Cell ◽  
Erythroid Progenitors ◽  
Globin Mrna ◽  
Erythroid Progenitor ◽  
Fetal Sheep ◽  
Gamma Globin

Cultures of peripheral blood or bone marrow erythroid progenitors display stimulated production of fetal hemoglobin. We investigated whether this stimulation is due to factors contained in the sera of the culture medium. Comparisons of gamma/gamma + beta biosynthetic ratios in erythroid colonies grown in fetal calf serum (FCS) or in charcoal treated FCS (C-FCS) showed that FCS-grown cells had significantly higher gamma/gamma + beta ratios. This increase in globin chain biosynthesis was reflected by an increase in relative amounts of steady- state gamma-globin mRNA. In contrast to its effect on adult cells, FCS failed to influence gamma-chain synthesis in fetal burst forming units- erythroid (BFU-E) colonies. There was a high correlation of gamma- globin expression in paired cultures done with C-FCS or fetal sheep serum. Dose-response experiments showed that the induction of gamma- globin expression is dependent on the concentration of FCS. These results indicate that FCS contains an activity that induces gamma- globin expression in adult erythroid progenitor cell cultures.

scholarly journals Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 918 ◽  
Author(s):  
Maria Karadjian ◽  
Anne-Sophie Senger ◽  
Christopher Essers ◽  
Sebastian Wilkesmann ◽  
Raban Heller ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Osteogenic Differentiation ◽  
Fetal Calf Serum ◽  
Culture Media ◽  
Human Platelet ◽  
Calf Serum ◽  
Calcium Content ◽  
Platelet Lysate ◽  
Protein Source ◽  
Human Platelet Lysate

Fetal calf serum (FCS) is frequently used as a growth factor and protein source in bone-marrow-derived mesenchymal stromal cell (BMSC) culture media, although it is a xenogenic product presenting multiple disadvantages including but not limited to ethical concerns. A promising alternative for FCS is human platelet lysate (hPL), which is produced out of human platelet concentrates and happens to be a stable and reliable protein source. In this study, we investigated the influence of hPL in an expansion medium (ESM) and an osteogenic differentiation medium (ODM) on the proliferation and osteogenic differentiation capacity of human BMSC. Therefore, we assessed population doublings during cell expansion, performed alizarin red staining to evaluate the calcium content in the extracellular matrix and determined the activity of alkaline phosphatase (ALP) as osteogenic differentiation correlates. The proliferation rate of BMSC cultured in ESM supplemented with hPL exceeded the proliferation rate of BMSC cultured in the presence of FCS. Furthermore, the calcium content and ALP activity was significantly higher in samples incubated in hPL-supplemented ODM, especially in the early phases of differentiation. Our results show that hPL can replace FCS as a protein supplier in cell culture media and does not negatively affect the osteogenic differentiation capacity of BMSC.

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.

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