scholarly journals Enrichment and characterization of thymus-repopulating cells in stroma-dependent cultures of rat bone marrow

1993 ◽  
Vol 104 (4) ◽  
pp. 1039-1048
Author(s):  
Z. Prakapas ◽  
M. Denoyelle ◽  
C. Dargemont ◽  
F.G. Kroese ◽  
J.P. Thiery ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Precursor Cells ◽  
Response Analysis ◽  
Mouse Bone Marrow ◽  
Low Density ◽  
Short Term ◽  
Fetal Mouse ◽  
Bone Marrow Precursor ◽  
Rat Bone

The bone marrow precursor cells seeding the thymus have been difficult to investigate using fresh bone marrow and in vivo thymus reconstitution assays. We have therefore designed a short-term bone marrow culture system allowing the study of thymus-repopulating cells in the marrow microenvironment. Low-density rat bone marrow cells were grown on pre-established mouse bone marrow stromal cell layers. Cocultured cells were maintained either under steroid-free conditions (Whitlock/Witte-type culture) or in the presence of 10(−7) M hydrocortisone (Dexter-type culture). After 3 days in vitro, the unanchored cell fractions were tested for their ability to colonize and repopulate fetal mouse thymic lobes in vitro. Both fresh low-density cells and Whitlock/Witte-type cultures, but not Dexter-type cultures, gave rise intrathymically to significant numbers of rat donor-type Thy-1.1high CD2+ CD5low CD43+ cells accounting for 50% to 90% of the organ-cultured cells at day 14. Repopulation of fetal mouse thymic lobes by rat Thy-1.1high cells could be used as a readout assay for initiation of thymopoiesis from bone marrow precursor cells, since 90% of the cells were CD3-/low and TCR alpha beta-/low and 15% of the cells co-expressed CD4 and CD8. Dose-response analysis showed that thymus repopulating cells were at least maintained, if not amplified during the 3-day culture period, leading to at least a 10-fold enrichment as compared to unfractionated bone marrow. Unlike fresh low-density cells before culture, short-term Whitlock/Witte-type cultures were depleted in myeloid-restricted precursor cells. In culture, the thymus-repopulating activity was predominantly associated with a 10% lymphoid cell subset which did not express the B-lineage-associated antigens revealed by HIS24 (the rat B220 equivalent) and HIS50 mAbs. We propose that unanchored thymus-repopulating cells enriched in Whitlock/Witte-type cultures may represent lymphoid-restricted, T-cell precursors of the bone marrow capable of emigrating and colonizing the thymus.

scholarly journals LncRNA Snhg6 regulates the differentiation of MDSCs by regulating the ubiquitination of EZH2

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Wei Lu ◽  
Fenghua Cao ◽  
Lili Feng ◽  
Ge Song ◽  
Yi Chang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Mrna Level ◽  
Suppressor Cells ◽  
Cell Group ◽  
Mouse Bone Marrow ◽  
Lewis Lung Cancer ◽  
Bone Marrow Precursor ◽  
Specific Regulation

AbstractMyeloid-derived suppressor cells (MDSCs) are derived from bone marrow progenitor cells commonly, which is a heterogeneous cell group composed of immature granulocytes, dendritic cells, macrophages and early undifferentiated bone marrow precursor cells. Its differentiation and immunosuppressive function are regulated by complex network signals, but the specific regulation mechanisms are not yet fully understood. In this study, we found that in mouse of Lewis lung cancer xenograft, long non-coding RNA Snhg6 (lncRNA Snhg6) was highly expressed in tumor-derived MDSCs compared with spleen-derived MDSCs. LncRNA Snhg6 facilitated the differentiation of CD11b+ Ly6G− Ly6Chigh monocytic MDSCs (Mo-MDSCs) rather than CD11b+ Ly6G+ Ly6Clow polymorphonuclear MDSCs (PMN-MDSCs), but did not affect the immunosuppressive function of MDSCs. Notably, lncRNA Snhg6 could inhibit the expression of EZH2 by ubiquitination pathway at protein level rather than mRNA level during the differentiation of mouse bone marrow cells into MDSCs in vitro. EZH2 may be an important factor in the regulation of lncRNA Snhg6 to promote the differentiation of Mo-MDSCs. So what we found may provide new ideas and targets for anti-tumor immunotherapy targeting MDSCs.

scholarly journals Regulation of megakaryopoiesis in long-term murine bone marrow cultures

Blood ◽  
1978 ◽  
Vol 51 (2) ◽  
pp. 245-255 ◽  
Author(s):  
N Williams ◽  
H Jackson ◽  
AP Sheridan ◽  
MJ Jr Murphy ◽  
A Elste ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Colony Formation ◽  
Leukemic Cell ◽  
Precursor Cells ◽  
Mouse Bone Marrow ◽  
Murine Bone Marrow ◽  
Marrow Cultures

Abstract Megakaryocytes and their precursor cells were sustained in mouse bone marrow suspension cultures for over 4–6 wk. Megakaryocyte precursor cells were detected by their capacity to form colonies of megakaryocytes in semisolid agar cultures. Colony formation was dependent on the presence of medium conditioned by a myelomonocytic leukemic cell line (WEHI-3CM). Megakaryocytes from the liquid and semisolid cultures were identified by cytoplasmic acetylcholine esterase and by ultrastructural analysis. The suspension medium from the bone marrow liquid cultures which sustained megakaryopoiesis was not directly acitive in stimulating megakaryocyte colony formation in the semisolid agar cultures, but potentiated the number of colonies detected when WEHI-3CM was present. Bone marrow-conditioned medium increased the sensitivity of megakaryocyte progenitor cells to the stimulus in WEHI-3CM. Addition of the activities present in the two sources produced a quantitative assay for the detection of mouse megakaryocyte progenitor cells. These studies showed: (1) that no inductive regulator of in vitro clones of megakaryocytes was present in the supernatants from the long-term marrow cultures and, (2) that at least two factors were necessary for the induction of megakaryocyte progenitors to proliferate and differentiate in semisolid cultures in vitro.

Short-term in vitro culture of purity and highly functional rat bone marrow-derived mast cells

2018 ◽  
Vol 54 (10) ◽  
pp. 705-714
Author(s):  
Tianyu Yu ◽  
Bin Liu ◽  
Zhigang He ◽  
Muqing Yang ◽  
Jian Song ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mast Cells ◽  
In Vitro Culture ◽  
Short Term ◽  
Rat Bone

Surface Electric Fields of Apatite Electret Promote Osteoblastic Responses

2012 ◽  
Vol 529-530 ◽  
pp. 357-360 ◽  
Author(s):  
Miho Nakamura ◽  
Akiko Nagai ◽  
Kimihiro Yamashita
Keyword(s):  
Bone Marrow ◽  
Quantitative Analysis ◽  
Electric Fields ◽  
Mtt Assay ◽  
Precursor Cells ◽  
Mouse Bone Marrow ◽  
Proliferation And Differentiation

The osteoblast behaviors on the biomaterial substrates are recognized to play a fundamental role in osteoconduction process. The purpose of this study was to evaluate the in vitro behaviors of osteoblasts cultured on electrically polarized hydroxyapatite (HA), having the enhanced osteobonding abilities. Osteoblasts derived from mouse bone marrow were seeded onto the polarized HA and investigated the proliferation and differentiation. The polarization had effects on the proliferation of osteoblast precursor cells based on the MTT assay. The acceleration was emerged as the early achievement to the confluence on the N-HA and P-HA. The quantitative analysis of the results of ALP and AR-S staining, the charges induced on the HA surface accelerated the differentiation from the osteoblast precursor cells to mature osteoblasts.

scholarly journals Regulation of megakaryopoiesis in long-term murine bone marrow cultures

Blood ◽  
1978 ◽  
Vol 51 (2) ◽  
pp. 245-255 ◽  
Author(s):  
N Williams ◽  
H Jackson ◽  
AP Sheridan ◽  
MJ Jr Murphy ◽  
A Elste ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Colony Formation ◽  
Leukemic Cell ◽  
Precursor Cells ◽  
Mouse Bone Marrow ◽  
Murine Bone Marrow ◽  
Marrow Cultures

Megakaryocytes and their precursor cells were sustained in mouse bone marrow suspension cultures for over 4–6 wk. Megakaryocyte precursor cells were detected by their capacity to form colonies of megakaryocytes in semisolid agar cultures. Colony formation was dependent on the presence of medium conditioned by a myelomonocytic leukemic cell line (WEHI-3CM). Megakaryocytes from the liquid and semisolid cultures were identified by cytoplasmic acetylcholine esterase and by ultrastructural analysis. The suspension medium from the bone marrow liquid cultures which sustained megakaryopoiesis was not directly acitive in stimulating megakaryocyte colony formation in the semisolid agar cultures, but potentiated the number of colonies detected when WEHI-3CM was present. Bone marrow-conditioned medium increased the sensitivity of megakaryocyte progenitor cells to the stimulus in WEHI-3CM. Addition of the activities present in the two sources produced a quantitative assay for the detection of mouse megakaryocyte progenitor cells. These studies showed: (1) that no inductive regulator of in vitro clones of megakaryocytes was present in the supernatants from the long-term marrow cultures and, (2) that at least two factors were necessary for the induction of megakaryocyte progenitors to proliferate and differentiate in semisolid cultures in vitro.

Resistance of Rat Bone Marrow Mesenchymal Stromal Precursor Cells to Anoxia In Vitro

2009 ◽  
Vol 148 (1) ◽  
pp. 148-151 ◽  
Author(s):  
E. B. Anokhina ◽  
L. B. Buravkova ◽  
S. V. Galchuk
Keyword(s):  
Bone Marrow ◽  
Precursor Cells ◽  
Stromal Precursor Cells ◽  
Rat Bone
Sign in / Sign up

Export Citation Format

Share Document