Immunocytochemistry of Blood and Bone Marrow Cells

Waele, Marc De; Beesley, Julian E.

doi:10.1016/b978-0-12-140407-9.50009-x

Cited by 4 publications

(3 citation statements)

References 86 publications

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“…Cytospin preparations were stained as described 42 with either rhodaminelabeled goat antimouse IgM or rhodamine-labeled goat antimouse IgG (Southern Biotechnology), and countersained with 4Ј,6-diamidino-2-phenylindole, dihydrochloride (DAPI; Molecular Probes, Eugene, OR). The slides were analyzed in a Leitz DMRD microscope (Leica, Wetzlar, Germany).…”

Section: Flow Cytometry and Immunofluorescence Microscopymentioning

confidence: 99%

Long-lived polyclonal B-cell lines derived from midgestation mouse embryo lymphohematopoietic progenitors reconstitute adult immunodeficient mice

Martı́nez-M.

Minguet²,

Gonzalo³

et al. 2001

Blood

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IntroductionThe scenario of mouse embryo lymphohematopoiesis has been transformed during recent years. Novel intraembryonic sites (paraaortic splanchnopleura/aorta-gonad-mesonephros region [P-Sp/ AGM], blood, omentum) and distinctive progenitors have been revealed in early, preliver midgestation periods (reviewed in Morales-Alcelay et al, 1 Melchers and Rolink, 2 and Keller et al 3 ). Stem cells giving rise to definitive lymphohematopoiesis are detected in para-aortic mesoderm, 4,5 as well as in yolk sac (YS), liver, blood, spleen and, finally, in bone marrow (BM) microenvironments. 6,7 Not only multipotential stem cells exist, but lineagespecific gene programs are also activated at the early ontogenic periods (days 9-12 postcoitum [dpc]); a limited process of B lymphopoiesis occurs, as it is revealed ex vivo by the detection of ckit ϩ AA4.1 ϩ CD19 ϩ cells, IgH DJ rearrangements, and the transcription of pre-B-specific genes (RAGs, VpreB, Ig␤, etc) in P-Sp/AGM, YS, and blood, and in liver at 11 to 12 dpc, [8][9][10] (De Andrés B et al, manuscript in preparation). These B-primed early progenitors maintain low cell numbers until days 12 to 13 of mouse gestation, and thereafter they grow exponentially. 9,[11][12][13][14][15] The early mouse embryo progenitors fully differentiate into B cells on embryo tissue grafting into severe combined immunodeficient (SCID) mice, 4 or after adoptive transfer experiments into conditioned newborn mice, 16 and they also mature in vitro on stromal cell/interleukin 7 (IL-7) cultures. 6 An efficient B lymphopoiesis relies on sequentially acting transcription factors (eg, Ikaros, Pax5, Id) that commit multipotential progenitors to the B-cell lineage, while restricting other cellular fates. [17][18][19] Supportive cytokines (especially IL-7) and interactions with stromal cells and components of the extracellular matrix, maintain viability and promote growth of early B-cell precursors (pro-B I cells) in inductive microenvironments. 20,21 The expression of pre-B-specific genes encoding for the recombinase enzymatic complex (RAGs, DNA-PK, Ku, others) and for the components of the surrogate light chain (SLC) (5, VpreB), is needed to proceed into differentiation. 22,23 The IgH chains that emerge from productive V H DJ H rearrangements and that are able to pair with the SLCs, are expressed in the membrane as pre-B-cell receptors (PreBCRs). [24][25][26] A second round of rearrangements takes place on the / loci of resting IgH ϩ pre-B cells to generate the light chains that are required to form a complete clonotypic H/L Ig receptor, which defines a B lymphocyte. The study of B-lineage cells in specific culture systems (stromal cells plus IL-7 and additional factors) has been highly instructive in elucidating the molecular steps of B-cell differentiation. 27 A drawback of the later protocols was the difficulty obtaining mature B cells in these cultures, leading to the suggestion of an IL-7-induced inhibition of late pre-B-cell differentiation. [27][28][29] The work reported here focuses on t...

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Section: Flow Cytometry and Immunofluorescence Microscopymentioning

confidence: 99%

Long-lived polyclonal B-cell lines derived from midgestation mouse embryo lymphohematopoietic progenitors reconstitute adult immunodeficient mice

Martı́nez-M.

Minguet²,

Gonzalo³

et al. 2001

Blood

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“…We used a method combining the cytochemical technique of selective elution of adult hemoglobin [12] and the imrnunogold-silverstaining method [13,14]. The RBCs from 40 ul of whole blood were washed three times with normal saline by centrifugation at 1,500g for 5 min.…”

Section: Laboratory Methodsmentioning

confidence: 99%

Maternal‐Fetal Hemorrhage: A Reappraisal

Brossard¹,

Pons

Jrad

et al. 1996

Vox Sanguinis

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“…We used a method combining the cytochemical technique of selec tive elution of adult hemoglobin [12] and the immunogold-silverstaining method [13,14]. The RBCs from 40 pi of whole blood were washed three times with normal saline by centrifugation at 1,500 g for 5 min.…”

Section: Laboratory Methodsmentioning

confidence: 99%

Maternal-Fetal Hemorrhage: A Reappraisal

Brossard¹,

Pons²,

Jrad³

et al. 1996

Vox Sanguinis

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To determine the frequency of maternal-fetal hemorrhage at or above 1 μl of maternal whole blood. Methods: Seventy-three mothers whose red blood cells bore an Rh antigen (Rh D, Rh c, Rh E) that was absent on red blood cells of their newborns were identified and a new cytological method, the Kleihauerimmunogold- silver-staining technique, was applied on the blood of their neonates to detect and quantify maternal red blood cells. Stringent precautions were taken to avoid contaminations of neonatal blood samples by adult red blood cells. Results: Maternal red blood cells were present in 3 newborns, a frequency of 4% (95% Cl: 1-11%), and the estimated volumes of hemorrhage were 0.8, 1.5, and 101 μl of maternal whole blood. No obstetric factor was clearly associated in this limited study with the occurrence of maternal-fetal hemorrhage. Conclusions: Mother-to-fetus microtransfusion greater than 1 μl is infrequent at or near delivery, and it may be observed after an uncomplicated pregnancy and vaginal delivery.

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Immunocytochemistry of Blood and Bone Marrow Cells

Cited by 4 publications

References 86 publications

Long-lived polyclonal B-cell lines derived from midgestation mouse embryo lymphohematopoietic progenitors reconstitute adult immunodeficient mice

Long-lived polyclonal B-cell lines derived from midgestation mouse embryo lymphohematopoietic progenitors reconstitute adult immunodeficient mice

Maternal‐Fetal Hemorrhage: A Reappraisal

Maternal-Fetal Hemorrhage: A Reappraisal

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