scholarly journals Mapping the Pairwise Choices Leading from Pluripotency to Human Bone, Heart, and Other Mesoderm Cell Types

Cell ◽  
2016 ◽  
Vol 166 (2) ◽  
pp. 451-467 ◽  
Author(s):  
Kyle M. Loh ◽  
Angela Chen ◽  
Pang Wei Koh ◽  
Tianda Z. Deng ◽  
Rahul Sinha ◽  
...  
Keyword(s):  
Cell Types ◽  
Human Bone ◽  
Mesoderm Cell

A role for cytoplasmic determinants in mesoderm patterning: cell-autonomous activation of the goosecoid and Xwnt-8 genes along the dorsoventral axis of early Xenopus embryos

Development ◽  
1994 ◽  
Vol 120 (5) ◽  
pp. 1191-1199 ◽  
Author(s):  
P. Lemaire ◽  
J.B. Gurdon
Keyword(s):  
Gene Activation ◽  
Cell Communication ◽  
Cell Types ◽  
Mesoderm Induction ◽  
Mesoderm Cell ◽  
Xenopus Embryos ◽  
Cell Dispersion ◽  
Embryo Axes ◽  
Alpha Actin ◽  
Cytoplasmic Determinants

Although an induction event is required for the formation of mesoderm in Xenopus embryos, it is not clear that this induction is wholly sufficient to give rise to a correctly patterned mesodermal layer. We have studied the expression of the two genes, goosecoid and Xwnt-8, in Xenopus gastrulae in which cell-cell communication, and therefore mesoderm induction, has been prevented by frequent cell dispersion. Although neither the early panmesodermal marker Xbra nor the muscle-specific alpha-actin gene were activated under these conditions, goosecoid and Xwnt-8 were activated in cells of dorsal and ventrolateral origin respectively, thus correctly reflecting their distribution during normal development. We also show that the spatial pattern of expression of these two genes along the animal-vegetal axis is similar in normal and in dissociated early gastrulae: goosecoid is mainly expressed in future mesoderm while the domain of expression of Xwnt-8 spans the mesoderm-endoderm boundary. These results show that, during the blastula and early gastrula stages, gene activation can be controlled cell-autonomously along both the animal-vegetal and dorsoventral embryo axes. This suggests that the inheritance of localised maternal cytoplasmic determinants is a key event for the patterning of mesoderm. We present a modified model of mesoderm formation in which the different mesoderm cell types are produced as a result of cooperation between induction-dependent and induction-independent immediate-early genes.

scholarly journals Dose-Response Tendon-Specific Markers Induction by Growth Differentiation Factor-5 in Human Bone Marrow and Umbilical Cord Mesenchymal Stem Cells

2020 ◽  
Vol 21 (16) ◽  
pp. 5905
Author(s):  
Maria Camilla Ciardulli ◽  
Luigi Marino ◽  
Erwin Pavel Lamparelli ◽  
Maurizio Guida ◽  
Nicholas Robert Forsyth ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Differentiation Factor ◽  
Anti Inflammatory ◽  
Growth Differentiation Factor 5

Mesenchymal stem cells derived from human bone marrow (hBM-MSCs) are utilized in tendon tissue-engineering protocols while extra-embryonic cord-derived, including from Wharton’s Jelly (hWJ-MSCs), are emerging as useful alternatives. To explore the tenogenic responsiveness of hBM-MSCs and hWJ-MSCs to human Growth Differentiation Factor 5 (hGDF-5) we supplemented each at doses of 1, 10, and 100 ng/mL of hGDF-5 and determined proliferation, morphology and time-dependent expression of tenogenic markers. We evaluated the expression of collagen types 1 (COL1A1) and 3 (COL3A1), Decorin (DCN), Scleraxis-A (SCX-A), Tenascin-C (TNC) and Tenomodulin (TNMD) noting the earliest and largest increase with 100 ng/mL. With 100 ng/mL, hBM-MSCs showed up-regulation of SCX-A (1.7-fold) at Day 1, TNC (1.3-fold) and TNMD (12-fold) at Day 8. hWJ-MSCs, at the same dose, showed up-regulation of COL1A1 (3-fold), DCN (2.7-fold), SCX-A (3.8-fold) and TNC (2.3-fold) after three days of culture. hWJ-MSCs also showed larger proliferation rate and marked aggregation into a tubular-shaped system at Day 7 (with 100 ng/mL of hGDF-5). Simultaneous to this, we explored the expression of pro-inflammatory (IL-6, TNF, IL-12A, IL-1β) and anti-inflammatory (IL-10, TGF-β1) cytokines across for both cell types. hBM-MSCs exhibited a better balance of pro-inflammatory and anti-inflammatory cytokines up-regulating IL-1β (11-fold) and IL-10 (10-fold) at Day 8; hWJ-MSCs, had a slight expression of IL-12A (1.5-fold), but a greater up-regulation of IL-10 (2.5-fold). Type 1 collagen and tenomodulin proteins, detected by immunofluorescence, confirming the greater protein expression when 100 ng/mL were supplemented. In the same conditions, both cell types showed specific alignment and shape modification with a length/width ratio increase, suggesting their response in activating tenogenic commitment events, and they both potential use in 3D in vitro tissue-engineering protocols.

scholarly journals Myofibroblastic stromal cells isolated from human bone marrow induce the proliferation of both early myeloid and B-lymphoid cells

Blood ◽  
1993 ◽  
Vol 82 (8) ◽  
pp. 2396-2405 ◽  
Author(s):  
I Moreau ◽  
V Duvert ◽  
C Caux ◽  
MC Galmiche ◽  
P Charbord ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Myeloid Cells ◽  
Cell Types ◽  
Lymphoid Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Present System ◽  
Normal Human ◽  
B Lymphoid

Abstract Normal human bone marrow stromal cells (BMSC) were isolated from Dexter- type long-term cultures according to their capacity to adhere to plastic and to their lack of hematopoietic antigens. The BMSC displayed a homogeneous appearance and a myofibroblastic phenotype in culture. The stromal cells (SC) were shown to support the proliferation of purified CD34+ hematopoietic progenitors and permitted us to maintain myeloid cells for several weeks in culture. In addition, the BMSC induced the proliferation of purified CD10+ s mu- fetal BM B-cell precursors (BCP). The capacity of the BMSC to induce the proliferation of early myeloid cells was shared by several other human fibroblastic- like cell types. In contrast, the BMSC were far superior to other adherent cells for induction of BCP proliferation. This capacity was largely mediated by endogenously produced interleukin-7 (IL-7), because it could be inhibited by anti-IL-7 antibody. In line with this finding, addition of IL-7 considerably enhanced BCP proliferation in cocultures with skin fibroblasts or synoviocytes. Thus, production of IL-7 appears to be a critical parameter that determines the ability of fibroblastic- like cells to induce BCP proliferation. Taken together, our data show that normal human myofibroblastic BMSC induce the proliferation of both early myeloid and B-lymphoid cells in the absence of accessory hematopoietic cells. The present system should constitute a model to study interactions between native human BM myofibroblastic stroma and various hematopoietic cell subsets.

scholarly journals Human eosinophil hematopoiesis studied in vitro by means of murine eosinophil differentiation factor (IL5): production of functionally active eosinophils from normal human bone marrow

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 646-651
Author(s):  
EJ Clutterbuck ◽  
CJ Sanderson
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Electron Microscopic ◽  
Human Eosinophil ◽  
Interleukin 5 ◽  
Differentiation Factor ◽  
Microscopic Features

The production of human eosinophils in vitro from normal bone marrow by using murine eosinophil differentiation factor (mEDF/interleukin 5) is described. Eosinophil production was selective and first detectable after 14 days and reached a peak between 21 and 35 days when they were the predominant cell type (41% to 89%). Until day 14, all the eosinophils were typical myelocytes, developing thereafter into metamyelocytes and mature cells. All cell types had characteristic light- and electron-microscopic features, apart from the absence of granules with crystalline cores. The eosinophils produced were readily recovered, and both immature myelocytes and mature cells were functionally active in an antibody-dependent, cell-mediated cytotoxicity assay. mEDF added into the assay enhanced the cytotoxicity but to a lower degree than previously reported for peripheral blood eosinophils, which suggests that they may be partially activated. The possibility that eosinophils could be deactivated was tested by removing mEDF from the culture medium. The eosinophils retained viability and functional activity, however, and showed no increased ability to be activated by mEDF for up to six days after removing the mEDF. The liquid culture of human bone marrow was shown to be an alternative assay for eosinophil differentiation factors to colony formation.

scholarly journals Human eosinophil hematopoiesis studied in vitro by means of murine eosinophil differentiation factor (IL5): production of functionally active eosinophils from normal human bone marrow

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 646-651 ◽  
Author(s):  
EJ Clutterbuck ◽  
CJ Sanderson
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Electron Microscopic ◽  
Human Eosinophil ◽  
Interleukin 5 ◽  
Differentiation Factor ◽  
Microscopic Features

Abstract The production of human eosinophils in vitro from normal bone marrow by using murine eosinophil differentiation factor (mEDF/interleukin 5) is described. Eosinophil production was selective and first detectable after 14 days and reached a peak between 21 and 35 days when they were the predominant cell type (41% to 89%). Until day 14, all the eosinophils were typical myelocytes, developing thereafter into metamyelocytes and mature cells. All cell types had characteristic light- and electron-microscopic features, apart from the absence of granules with crystalline cores. The eosinophils produced were readily recovered, and both immature myelocytes and mature cells were functionally active in an antibody-dependent, cell-mediated cytotoxicity assay. mEDF added into the assay enhanced the cytotoxicity but to a lower degree than previously reported for peripheral blood eosinophils, which suggests that they may be partially activated. The possibility that eosinophils could be deactivated was tested by removing mEDF from the culture medium. The eosinophils retained viability and functional activity, however, and showed no increased ability to be activated by mEDF for up to six days after removing the mEDF. The liquid culture of human bone marrow was shown to be an alternative assay for eosinophil differentiation factors to colony formation.

Hematopoiesis in Three-Dimensions: Tissue Architecture of Murine and Human Bone Marrow Visualized to a Depth of 100 Micron by Confocal Microscopy

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4781-4781
Author(s):  
Tomoiku Takaku ◽  
Daniela Malide ◽  
Ning Zhi ◽  
Rodrigo Calado ◽  
Jichun Chen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Extracellular Matrix ◽  
Laser Scanning ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Bone Marrow Tissue ◽  
Murine Bone Marrow ◽  
3 Dimensional ◽  
Whole Mount

Abstract Three-dimensional (3D) reconstruction of organs and tissues is a powerful tool to establish anatomical and functional relationships of microscopic structures. We developed whole-mount tissue processing methods for 3D in situ visualization of murine and human bone marrow; our methods are compatible with fluorescent labeling of different cell types and other structures of interest in the tissue microenvironment. The major technical problems addressed were the conditions for tissue fixation in the absence of permeabilization and sectioning; antibody penetration and binding; and the acquisition of high quality images by adequate laser scanning confocal microscope. For murine bone marrow, the sternum was bisected sagitally; for human tissue, 2–3 mm fragments of core biopsies were utilized. Bone marrow tissue and cells were exposed to fluorescence labeled nucleic acid dyes and antibodies, with or without prior chemical fixation. Single and double labeling of cells was feasible with combinations of various antibodies and direct and indirect immunofluorescent techniques. In some experiments, cells were visualized from transgenic mice with cell populations expressing green fluorescence protein (GFP). Series of two dimensional (xy) images 600 μm × 600 μm were collected along the z-axis at 5 μm z-intervals to depths of 60–100 μm using a Zeiss LSM 510 confocal microscope. Two dimensional images were assembled to reconstruct 3-dimensional volumes by Bitplane’s Imaris 3D computer software. Antigenicity was preserved, allowing simultaneous labeling of cell types and structures by immunohistochemistry or nuclear dyes. Different hematopoietic cell types as well as blood vessels, adipose cells, and extracellular matrix were visualized in complex 3-dimensional organization of intact bone marrow tissue revealing unknown features of multicellular architecture. Normal murine bone marrow, after brief fixation formaldehyde, is shown in Figure A. Rat anti-mouse basement-membrane monoclonal antibody (MAb) and fluorescent isothiocyanate (FITC)-labeled donkey anti-rat monoclonal antibody were used to visualize the extracellular matrix and micro-vessels (appearing green). Allophycocyanin (APC)-labeled rat anti-mouse CD45R cells permitted visualization of B lymphocytes (red). 4’,6-diamidino-2-phenylindole(DAPI) stained all nuclei (blue). Nests of lymphocytes appeared encased by extracellular matrix, fed by microvessels running from the bone edge. An example of the architecture of a human hematologic malignancy is shown in figure B, from a marrow biopsy of a patient with multiple myeloma prior to therapy. Mouse anti-human CD20 MAb and FITC-labeled donkey anti-mouse IgG were used to visualize mature B cells (green). APC-conjugated mouse anti-human CD38 MAb identified plasma cells (red). DAPI stained nuclei (blue). The large tumor cells appeared in unevenly distributed cell clumps. In mouse experiments, (not illustrated), marrow cells were easily observed in animals in which GFP was driven by the ubiquitin-C promoter. In humans (also not illustrated), we observed malignant cell populations stained with appropriate lineage-specific antibodies in patients with leukemia and compared CD34 cell numbers in normal with aplastic bone marrow. Confocal laser scanning microscopy, a powerful technique to generate serial sections of whole-mount tissue and their digital reassembly into virtual 3-dimensional structures, has been readily adapted to examination of murine and human bone marrow. The wide variety of MAbs available for specific antigens in combination with this imaging method should aid in conceptualizing microanatomical relationships among hematopoietic cells, stroma, blood vessels, and extracellular matrix in normal and diseased bone marrow. Figure Figure

Continuous human bone marrow culture: Ia antigen characterization of probable pluripotential stem cells

Blood ◽  
1980 ◽  
Vol 55 (4) ◽  
pp. 682-690 ◽  
Author(s):  
MA Moore ◽  
HE Broxmeyer ◽  
AP Sheridan ◽  
PA Meyers ◽  
N Jacobsen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Progenitor Cell ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Cell Type ◽  
Granulocytic Colony ◽  
A Cell

Abstract The presence of Ia-like antigens on human CFU-C and BFU-e is confirmed and a cell type that lacked immediate capacity for granulocytic colony formation but generated CFU-c after brief incubation in simple suspension culture is identified. This pre-CFU-c, and its immediate progeny, was extremely sensitive to killing by anti-Ia serum with complement. In contrast, anti-Ia serum plus complement treatment of human bone marrow, while eliminating 93%-97% of all CFU-c and BFU-e, did not prevent the rapid regeneration of these progenitor cells and their production for some weeks under the conditions of continuous marrow culture. These studies suggest that the human equivalent of the pluripotential stem cell can replicate for some weeks in culture and generate committed progenitors, such as CFU-c and BFU-e. Furthermore, it would appear that Ia-like antigen is absent on the pluripotential stem cell, is rapidly gained as commitment to the various progenitor cell types occur, and is subsequently lost as these latter undergo differentiation within the marrow.

scholarly journals Human immunodeficiency virus infection of human bone marrow stromal fibroblasts

Blood ◽  
1990 ◽  
Vol 76 (2) ◽  
pp. 317-322 ◽  
Author(s):  
DT Scadden ◽  
M Zeira ◽  
A Woon ◽  
Z Wang ◽  
L Schieve ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Immunodeficiency Virus ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Stromal Fibroblast ◽  
Stromal Fibroblasts ◽  
Fibroblast Line ◽  
Immunodeficiency Virus

Abstract The human immunodeficiency virus (HIV) preferentially infects CD4 positive T cells and monocytes. Other human cell types have been reported to be infectable with HIV, including cells of mesenchymal origin. In this report, we show that both primary human bone marrow stromal fibroblasts and an immortalized human stromal fibroblast line are susceptible to HIV infection. These cells are capable of passing HIV to cells of lymphoid or myeloid lineage, and may thereby act as a reservoir of virus. This in vitro system may be a useful model for assessing the pathophysiology of hematopoietic dysfunction in AIDS patients.

Interleukin-6 is not involved in the interleukin-1-induced production of colony-stimulating factors by human bone marrow stromal cells and fibroblasts

Blood ◽  
1989 ◽  
Vol 74 (8) ◽  
pp. 2619-2623
Author(s):  
MR Schaafsma ◽  
WE Fibbe ◽  
J Van Damme ◽  
N Duinkerken ◽  
P Ralph ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Interleukin 6 ◽  
Interleukin 1 ◽  
Human Fibroblasts ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Colony Stimulating Factors ◽  
Csf Production

Interleukin-6 (IL-6) is a multifunctional cytokine that plays a role in regulation of hematopoiesis. Because IL-6 is coinduced with colony- stimulating factors (CSFs) by various cell types in response to stimulation with IL-1, we investigated whether IL-6 is involved in the IL-1-induced production of CSF by human bone marrow (BM) cells in long- term culture or human fibroblasts. We showed that IL-6 does not induce CSF production by these cells. Neither addition of exogenous IL-6 nor neutralization of endogenous production of IL-6 by an anti-IL-6 monoclonal antibody (MoAb) diminished the IL-1-induced colony- stimulating activity (CSA), indicating that IL-6 did not act synergistically with IL-1. Finally, IL-6 did not influence the kinetics of IL-1-induced CSA production by human fibroblasts. We conclude that IL-6, either alone or in combination with IL-1, does not induce CSF production by human BM stromal cells or fibroblasts.

scholarly journals Interleukin-6 is not involved in the interleukin-1-induced production of colony-stimulating factors by human bone marrow stromal cells and fibroblasts

Blood ◽  
1989 ◽  
Vol 74 (8) ◽  
pp. 2619-2623 ◽  
Author(s):  
MR Schaafsma ◽  
WE Fibbe ◽  
J Van Damme ◽  
N Duinkerken ◽  
P Ralph ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Interleukin 6 ◽  
Interleukin 1 ◽  
Human Fibroblasts ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Colony Stimulating Factors ◽  
Csf Production

Abstract Interleukin-6 (IL-6) is a multifunctional cytokine that plays a role in regulation of hematopoiesis. Because IL-6 is coinduced with colony- stimulating factors (CSFs) by various cell types in response to stimulation with IL-1, we investigated whether IL-6 is involved in the IL-1-induced production of CSF by human bone marrow (BM) cells in long- term culture or human fibroblasts. We showed that IL-6 does not induce CSF production by these cells. Neither addition of exogenous IL-6 nor neutralization of endogenous production of IL-6 by an anti-IL-6 monoclonal antibody (MoAb) diminished the IL-1-induced colony- stimulating activity (CSA), indicating that IL-6 did not act synergistically with IL-1. Finally, IL-6 did not influence the kinetics of IL-1-induced CSA production by human fibroblasts. We conclude that IL-6, either alone or in combination with IL-1, does not induce CSF production by human BM stromal cells or fibroblasts.

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