Migration and morphology changes of bone marrow cells in ischemic and peri-ischemic areas of the mouse brain demonstrated by chronic in vivo imaging

Abstract While hematopoietic engraftment kinetics are well appreciated after lethal irradiation in the mouse, most observations have been limited to blood samples or terminal examination of marrow or spleen. The development of non-invasive bioluminescence in vivo imaging technology allows a dynamic picture of engraftment and clonal expansion to be defined. We have extended this technology to the process of drug resistance gene therapy. We hypothesized that drug selection would profoundly affect the extent and dynamics of hematopoietic stem cells (HSC) engraftment and clonal expansion after lentiviral mediated gene transfer of the P140KMGMT gene into murine HSC. In previous studies, we have shown that P140KMGMT gene containing retroviral and lentiviral transduced bone marrow cells provided significant protection against chemotherapeutic drugs BCNU and TMZ given with BG (O6-Benzylguanine), in vitro and in vivo. We generated a bicistronic lentiviral vector containing P140KMGMT gene and firefly luciferase gene linked by 2A sequence of FMDV(Foot-and-Mouth Disease Virus), which will cleave itself during ribosomal translation. Whole bone marrow cells was collected from BALB/c mice 4 days after 5-FU treatment and transduced with P140KMGMT-luc lentiviruses at MOI of 1.4. Transduced bone marrow cells were transplanted into lethally irradiated or non-myeloablated syngeneic recipient mice at different cell numbers. Initial bioluminescent signal emerged 6–8 days after transplantation in both lethally irradiated and non-myeloablated recipients. The onset of bioluminescent foci after transplantation occurred in a cell dose dependent manner. The initial signal emitted predominantly from bone marrow, especially femurs, humeri and vertebrae during the early stage of clonal expansion. Intense signal appeared in spleen at days 12–14 and became weaker or even disappeared by days 20–28. Clonal expansion and engraftment greatly increased after a single course of BG+TMZ treatment and initiated strong hematopoiesis in non-myeloablated recipients. Total body bioluminescence intensity of drug treated mice increased 24 fold and 7 fold compared to non-treated mice in both non-myeloablated and lethally irradiated recipients, respectively. A transient phase suggesting migration through the lymphatic system and in the spleen occurred in most mice and was exacerbated by drug selection, but this was less clear in lethally irradiated mice, where engraftment was more confined to the marrow spaces. Bioluminescence in vivo imaging reveals active migration between the bone marrow and the spleen during hematopoiesis. Drug selection has a significant impact on the patterns of engraftment and clonal expansion of HSC and progenitor cells after transplantation.

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In vivo imaging with cellular resolution of bone marrow cells transplanted into the ischemic brain of a mouse

NeuroImage ◽

10.1016/j.neuroimage.2006.01.019 ◽

2006 ◽

Vol 31 (3) ◽

pp. 958-967 ◽

Cited By ~ 11

Author(s):

Alexy Tran Dinh ◽

Nathalie Kubis ◽

Yutaka Tomita ◽

Bartosz Karaszewski ◽

Yolande Calando ◽

...

Keyword(s):

Bone Marrow ◽

In Vivo Imaging ◽

Bone Marrow Cells ◽

Ischemic Brain ◽

Cellular Resolution ◽

Marrow Cells

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CD34+ human marrow cells that express low levels of Kit protein are enriched for long-term marrow-engrafting cells

Blood ◽

10.1182/blood.v87.10.4136.bloodjournal87104136 ◽

1996 ◽

Vol 87 (10) ◽

pp. 4136-4142 ◽

Cited By ~ 113

Author(s):

I Kawashima ◽

ED Zanjani ◽

G Almaida-Porada ◽

AW Flake ◽

H Zeng ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Marrow Cell ◽

In Utero ◽

Fetal Sheep ◽

Hematopoietic Stem ◽

Show Evidence ◽

Marrow Cells

Using in utero transplantation into fetal sheep, we examined the capability of human bone marrow CD34+ cells fractionated based on Kit protein expression to provide long-term in vivo engraftment. Twelve hundred to 5,000 CD34+ Kit-, CD34+ Kit(low), and CD34+ Kit(high) cells were injected into a total of 14 preimmune fetal sheep recipients using the amniotic bubble technique. Six fetuses were killed in utero 1.5 months after bone marrow cell transplantation. Two fetuses receiving CD34+ Kit(low) cells showed signs of engraftment according to analysis of CD45+ cells in their bone marrow cells and karyotype studies of the colonies grown in methylcellulose culture. In contrast, two fetuses receiving CD34+ Kit(high) cells and two fetuses receiving CD34+ Kit- cells failed to show evidence of significant engraftment. Two fetuses were absorbed. A total of six fetuses receiving different cell populations were allowed to proceed to term, and the newborn sheep were serially examined for the presence of chimerism. Again, only the two sheep receiving CD34+ Kit(low) cells exhibited signs of engraftment upon serial examination. Earlier in studies of murine hematopoiesis, we have shown stage-specific changes in Kit expression by the progenitors. The studies of human cells reported here are in agreement with observations in mice, and indicate that human hematopoietic stem cells are enriched in the Kit(low) population.

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Distribution of sister chromatid exchanges on the mouse chromosomes in vivo with reference to the replication properties of the X chromosome

Canadian Journal of Genetics and Cytology ◽

10.1139/g84-026 ◽

1984 ◽

Vol 26 (2) ◽

pp. 152-157

Author(s):

S. M. Singh ◽

D. L. Reimer

Keyword(s):

Bone Marrow ◽

X Chromosome ◽

Sister Chromatid ◽

Bone Marrow Cells ◽

Relative Length ◽

Chromosome Length ◽

Sister Chromatid Exchanges ◽

Chromatid Exchanges ◽

Marrow Cells

Frequency of sister chromatid exchanges (SCE) were recorded separately for different chromosomes from bone marrow cells of female mice of the two genetic strains (C3H/S and C57BL/6J). SCEs were evaluated following different doses of 5-bromo-2′deoxyuridine (BrdU) as nine hourly i.p. injections. The SCE per cell increased with increasing BrdU doses which was slightly higher in C3H/S than in the C57BL/6J. SCEs per cell were variable at every treatment – strain combination, possibly reflecting the heterogeneous nature of the bone marrow cells. In general, there is a positive correlation between SCE per chromosome and the relative chromosome length. Total SCEs on one of the large chromosomes (most likely the X chromosome), however, are significantly higher than expected on the basis of relative length alone. Most of this increase is attributable to one of the homologues of this chromosome, which is not in synchrony with the rest of the chromosomes and may represent the late-replicating X. These results when viewed in the light of replication properties of the heterochromatinized X, suggest a direct involvement of DNA replication in SCE formation and may argue against the replication point as the sole site for the SCEs.Key words: sister chromatid exchange, BrdU, recombination, replication, X chromosome.

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Mutagenicity assay in Salmonella and in vivo sister chromatid exchange in bone marrow cells of mice for four pyrazolone derivatives

Mutation Research/Genetic Toxicology and Environmental Mutagenesis ◽

10.1016/s1383-5718(98)00138-7 ◽

1998 ◽

Vol 420 (1-3) ◽

pp. 15-25 ◽

Cited By ~ 18

Author(s):

A.K. Giri ◽

A. Mukhopadhyay

Keyword(s):

Bone Marrow ◽

Sister Chromatid ◽

Sister Chromatid Exchange ◽

Bone Marrow Cells ◽

Pyrazolone Derivatives ◽

Mutagenicity Assay ◽

Marrow Cells

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AML cells are differentially sensitive to chemotherapy treatment in a human xenograft model

Blood ◽

10.1182/blood-2012-10-464677 ◽

2013 ◽

Vol 121 (12) ◽

pp. e90-e97 ◽

Cited By ~ 66

Author(s):

Mark Wunderlich ◽

Benjamin Mizukawa ◽

Fu-Sheng Chou ◽

Christina Sexton ◽

Mahesh Shrestha ◽

...

Keyword(s):

Bone Marrow ◽

Induction Therapy ◽

Bone Marrow Cells ◽

Xenograft Model ◽

Chemotherapy Treatment ◽

Key Points ◽

Increased Sensitivity ◽

Total Bone Marrow ◽

Marrow Cells

Key Points A relevant xenograft chemotherapy model was developed by using standard AML induction therapy drugs and primary human AML patient samples. Human AML cells show significantly increased sensitivity to in vivo chemotherapy treatment compared with murine LSK and total bone marrow cells.

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