In Vitro Development of Bone-Marrow-Derived Macrophages Influence of Mouse Genotype on Response to Colony-Stimulating Factors and Autocrine Interferon Induction

1989 ◽  
Vol 30 (6) ◽  
pp. 731-740 ◽  
Author(s):  
A. KRUSE ◽  
H. KIRCHNER ◽  
R. ZAWATZKY ◽  
I. DOMKE-OPITZ
Keyword(s):  
Bone Marrow ◽  
Colony Stimulating Factors ◽  
In Vitro Development ◽  
Mouse Genotype ◽  
Interferon Induction ◽  
Vitro Development

Bloodstream Progenitor-Cell Traffic In Primary Myelofibrosis Reveals Cell Subsets Showing Some Features Of Very-Small Embryonnic-Like Stem Cells (VSELs), Along With Endothelial (PEC), Mesenchymal (MPC) and Hematopoietic (HPC) Progenitor Cells

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5265-5265 ◽  
Author(s):  
Mario Ojeda-Uribe ◽  
Hanna Sovalat ◽  
Laura Jung ◽  
Christophe Desterke ◽  
Sylvie Thiebault ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Hot Spot ◽  
Embryonic Stem ◽  
Primary Myelofibrosis ◽  
Rt Pcr ◽  
In Vitro Development ◽  
Oct4 Expression ◽  
Vitro Development

Abstract Introduction Primary myelofibrosis (PMF) is accompanied by an increase in the bloodstream circulation of some adult progenitor cells. Extramedullary hematopoiesis observed in this setting might remind some features related to foetal hematopoiesis. Material and methods We looked for evidence in favour of this hypothesis in blood samples of a small cohort of untreated patients with PMF (4 pre-fibrotic (PF) and 4 fibrotic (F), defined according to the WHO and Thiele's histopathology score (Blood, 2011)). Patient baseline characteristics are shown below. We performed a) flow-cytometric analysis for cell subsets related to VSEL, PEC, MPC, HPC; b) RT-PCR for embryonic transcriptional factors NANOG, OCT4, SOX2, LIN28 from MNC fraction (positive control hES, negative control CPRE2 c) in-vitro development of embryonic stem like cells (ESlC) under specific culture conditions. In addition we looked for SRSF2 mutations in order to better characterize PMF stages. Results As expected we detected high numbers of circulating CD34+ cells (HPC) (mean 233083±307148/ml (range 4600-783000), with similar numbers in PF- (231125±289553/ml) and F-PMF (235040 ±369156/ml). We were able to detect small numbers of the following cell subsets related to VSEL (size 2-4m) (Fig 1) Lin-/CD45-/CD34+ (mean 124±239/ml), Lin-/CD45-/CD133+ (mean 1178±971/ml), Lin-/CD45-/CXCR4+ (mean 1572±1622/ml). Lin-/CD45-CD34+AC133+CXCR4+ cells were detected in 6 of 8 patients (mean 186±375/ml) with F-PMF patients showing higher numbers (279±416/ml) than PF-PMF (63±71/ml). NANOG and OCT4 expression was detected by RT-PCR in all the patients tested. Mean OCT4 expression was about 50% the level of hES, but F-PMF showed higher levels. NANOG expression was similar to that of hES, whereas Sox2 and Lin28 were not expressed in most patients. We failed to observe the in-vitro development of ESlC in the 2 tested patients. PEC (Lin-/CD45-CD34+AC133+KDR+) were detected in all the PF-PMF (185±332/ml) and in 1 of 4 F-PMF (mean 9±18/ml). MPC (Lin-/CD45-CD90+CD105+) were detected in higher numbers in PF-PMF (mean 413±528/ml) than in F-PMF (mean 157±216/ml). We were not able to detect mutations in the hot spot of SRSF2 (codons 93,94,95). Conclusions Small numbers of cell subsets displaying morphologic and immunophenotypic features of VSELs were detected in PMF patients. However, we are not able to define these as fully specific VSELs according to previous works that defined them (Kucia, Leukemia 2006). Interestingly Lin-/CD45-CD34+AC133+CXCR4+ cells were observed in higher numbers in F-PMF, supporting in part our hypothesis that PMF evolution can be associated to the recruitment and circulation of some primitive progenitors (dormant in the adult life) as it can be observed during the foetal period. This recruitment also involves HPC. Moreover although all patients expressed OCT4 and NANOG, OCT4 expression was higher in F-PMF. As expected PEC circulate in higher numbers in PF-PMF compared to F-PMF. Interestingly both F-PMF and PF-PMF were associated to the circulation of significant numbers of MPC but higher numbers observed in PF-MFP might be interpreted either as a necessary recruitment to establish extra-medullary stroma or due to the exit from bone marrow of highly proliferative MPC. Whether all these different circulating progenitor cells, are clonally or not clonally related to the PMF pathogenesis or to unspecific mobilisation secondary to bone marrow microenvironment injury cannot be determined from these preliminary results. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Delineation of precursors in murine spleen that develop in contact with splenic endothelium to give novel dendritic-like cells

Blood ◽  
2010 ◽  
Vol 115 (18) ◽  
pp. 3678-3685 ◽  
Author(s):  
Jonathan K. H. Tan ◽  
Pravin Periasamy ◽  
Helen C. O'Neill
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Culture System ◽  
In Vitro Development ◽  
Cell Lineages ◽  
Mhc Ii ◽  
Murine Spleen ◽  
Vitro Development

Abstract Hematopoietic cell lineages are best described in terms of distinct progenitors with limited differentiative capacity. To distinguish cell lineages, it is necessary to define progenitors and induce their differentiation in vitro. We previously reported in vitro development of immature dendritic-like cells (DCs) in long-term cultures (LTCs) of murine spleen, and in cocultures of spleen or bone marrow (BM) over splenic endothelial cell lines derived from LTCs. Cells produced are phenotypically distinct CD11bhiCD11cloCD8−MHC-II− cells, tentatively named L-DCs. Here we delineate L-DC progenitors as different from known DC progenitors in BM and DC precursors in spleen. The progenitor is contained within the lineage-negative (Lin)−c-kit+ subset in neonatal and adult spleen. This subset has multipotential reconstituting ability in mice. In neonatal spleen, the progenitor is further enriched within the c-kitlo and CD34+ subsets of Lin−c-kit+ cells. These cells seed cocultures of splenic endothelial cells, differentiating to give L-DCs that can activate T cells. L-DC progenitors are distinguishable from described splenic CD11clo DC precursors and from Fms-like tyrosine kinase 3+ DC progenitors in BM. Overall, this study confirms that LTCs are a physiologically relevant culture system for in vitro development of a novel DC type from spleen progenitors.

scholarly journals Imatinib inhibits the in vitro development of the monocyte/macrophage lineage from normal human bone marrow progenitors

Leukemia ◽  
2003 ◽  
Vol 17 (9) ◽  
pp. 1713-1721 ◽  
Author(s):  
A L Dewar ◽  
R M Domaschenz ◽  
K V Doherty ◽  
T P Hughes ◽  
A B Lyons
Keyword(s):  
Bone Marrow ◽  
Human Bone ◽  
Human Bone Marrow ◽  
In Vitro Development ◽  
Normal Human ◽  
Vitro Development ◽  
Normal Human Bone Marrow ◽  
Macrophage Lineage

In vitro development of human triploid zygotes reconstructed by pronuclear transfer

2002 ◽  
Vol 78 ◽  
pp. S180-S181
Author(s):  
John Zhang ◽  
Yi Ming Shu ◽  
Lewis C Krey ◽  
Hui Liu ◽  
Guang Lun Zhuang ◽  
...  
Keyword(s):  
In Vitro Development ◽  
Pronuclear Transfer ◽  
Vitro Development

Gallic acid promotes the in vitro development of sheep secondary isolated follicles involving the phosphatidylinositol 3-kinase pathway

2021 ◽  
pp. 106767
Author(s):  
Gizele A.L. Silva ◽  
Luana B. Araújo ◽  
Larissa C.R. Silva ◽  
Bruna B. Gouveia ◽  
Ricássio S. Barberino ◽  
...  
Keyword(s):  
Gallic Acid ◽  
Phosphatidylinositol 3 Kinase ◽  
In Vitro Development ◽  
Vitro Development ◽  
Kinase Pathway ◽  
Phosphatidylinositol 3

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

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