Intra-bone Marrow Transplant (IBMT) of Cord Blood (CB) Cells: A Transplant Approach that Tries to Optimize Seeding Efficiency and Trafficking of Hematopoietic Stem Cells (HSCs)

2014 ◽  
pp. 203-210
Author(s):  
Francesco Frassoni ◽  
Francesca Bonifazi ◽  
Marina Podestà ◽  
Giuseppe Bandini ◽  
Daniela Cilloni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Bone Marrow Transplant ◽  
Marrow Transplant ◽  
Hematopoietic Stem ◽  
Seeding Efficiency

scholarly journals Multimodal Tracking of Hematopoietic Stem Cells from Young and Old Mice Labeled with Magnetic–Fluorescent Nanoparticles and Their Grafting by Bioluminescence in a Bone Marrow Transplant Model

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 752
Author(s):  
Fernando A. Oliveira ◽  
Mariana P. Nucci ◽  
Javier B. Mamani ◽  
Arielly H. Alves ◽  
Gabriel N. A. Rego ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Bone Marrow Transplant ◽  
Marrow Transplant ◽  
Hematopoietic Stem ◽  
Icp Ms ◽  
Old Mice

This study proposes an innovative way to evaluate the homing and tracking of hematopoietic stem cells from young and old mice labeled with SPIONNIRF-Rh conjugated with two types of fluorophores (NIRF and Rhodamine), and their grafting by bioluminescence (BLI) in a bone marrow transplant (BMT) model. In an in vitro study, we isolated bone marrow mononuclear cells (BM-MNC) from young and old mice, and analyzed the physical–chemical characteristics of SPIONNIRF-Rh, their internalization, cell viability, and the iron quantification by NIRF, ICP-MS, and MRI. The in vivo study was performed in a BMT model to evaluate the homing, tracking, and grafting of young and old BM-MNC labeled with SPIONNIRF-Rh by NIRF and BLI, as well as the hematological reconstitution for 120 days. 5FU influenced the number of cells isolated mainly in young cells. SPIONNIRF-Rh had adequate characteristics for efficient internalization into BM-MNC. The iron load quantification by NIRF, ICP-MS, and MRI was in the order of 104 SPIONNIRF-Rh/BM-MNC. In the in vivo study, the acute NIRF evaluation showed higher signal intensity in the spinal cord and abdominal region, and the BLI evaluation allowed follow-up (11–120 days), achieving a peak of intensity at 30 days, which remained stable around 108 photons/s until the end. The hematologic evaluation showed similar behavior until 30 days and the histological results confirm that iron is present in almost all tissue evaluated. Our results on BM-MNC homing and tracking in the BMT model did not show a difference in migration or grafting of cells from young or old mice, with the hemogram analysis trending to differentiation towards the myeloid lineage in mice that received cells from old animals. The cell homing by NIRF and long term cell follow-up by BLI highlighted the relevance of the multimodal nanoparticles and combined techniques for evaluation.

scholarly journals Searching for unrelated donor hematopoietic stem cells: Availability and speed of umbilical cord blood versus bone marrow

2002 ◽  
Vol 8 (5) ◽  
pp. 257-260 ◽  
Author(s):  
Juliet N Barker ◽  
Timothy P Krepski ◽  
Todd E DeFor ◽  
Stella M Davies ◽  
John E Wagner ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Unrelated Donor ◽  
Hematopoietic Stem

Mesenchymal Stem Cells from the Wharton’s Jelly of Umbilical Cord Segments Support the Maintenance of Long Term Culture-Initiating Cells from Cord Blood.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3650-3650
Author(s):  
Kent W. Christopherson ◽  
Tiki Bakhshi ◽  
Shamanique Bodie ◽  
Shannon Kidd ◽  
Ryan Zabriskie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Blood Cells ◽  
Hematopoietic Stem ◽  
Cord Blood Cells

Abstract Hematopoietic Stem Cells (HSC) are routinely obtained from bone marrow, mobilized peripheral blood, and umbilical Cord Blood. Traditionally, adult bone marrow has been utilized as a source of Mesenchymal Stem Cells (MSC). Bone marrow derived MSC (BM-MSC) have previously been shown to maintain the growth of HSC obtained from cord blood and have been utilized for cord blood expansion purposes. However, the use of a mismatched BM-MSC feeder stromal layer to support the long term culture of cord blood HSC is not ideal for transplant purposes. The isolation of MSC from a novel source, the Wharton’s Jelly of Umbilical Cord segments, was recently reported (Romanov Y, et al. Stem Cells.2003; 21: 105–110) (Lee O, et al. Blood.2004; 103: 1669–1675). We therefore hypothesized that Umbilical Cord derived MSC (UC-MSC) have the ability to support the long term growth of cord blood derived HSC similar to that previously reported for BM-MSC. To test this hypothesis, MSC were isolated from the Wharton’s Jelly of Umbilical Cord segments and defined morphologically and by cell surface markers. UC-MSC were then tested for their ability to support the growth of pooled CD34+ cord blood cells in long term culture - initiating cell (LTC-IC) assays as compared to BM-MSC. We observed that like BM-MSC, CB-MSC express a defined set of cell surface markers. By flow cytometry we determined that that both UC-MSC and BM-MSC are positive for CD29, CD44, CD73, CD90, CD105, CD166, HLA-A and negative for CD45, CD34, CD38, CD117, HLA-DR expression. Utilizing Mitomycin C treated (200 μM, 15 min.) UC-MSC from multiple donors as a feeder layer we observed that UC-MSC have the ability to support the maintenance of long term hematopoiesis during the LTC-IC assay. Specifically, UC-MSC isolated from separate umbilical cord donors support the growth of 69.6±11.9 (1A), 31.7±3.9 (2B), 67.0±13.5 (3A), and 38.5±13.7 (3B) colony forming cells (CFC) per 1×104 CD34+ cord blood cells as compared to 64.0±4.2 CFC per 1×104 CD34+ cord blood cells supported by BM-MSC (Mean±SEM, N=4 separate segments from three different donors). Thus, Umbilical Cord derived Mesenchymal Stem Cells, a recently described novel source of MSC, have the ability to support long term maintenance of Hematopoietic Stem Cells, as defined by the LTC-IC assay. These results may have potential therapeutic application with respect to ex vivo stem cell expansion of Cord Blood Hematopoietic Stem Cells utilizing a Mesenchymal Stem Cell stromal layer. In addition, these data suggest the possibility of co-transplantation of matched Mesenchymal and Hematopoietic Stem Cells from the same umbilical cord and cord blood donor respectively. Lastly, these results describe a novel model system for the future study of the interaction between Cord Blood Hematopoietic Stem Cells and the appropriate supportive microenvironment represented by the Umbilical Cord - Mesenchymal Stem Cells.

Donor-Derived Hematopoietic Stem Cells Repair the Bone Marrow Sinusoids Following Bone Marrow Transplant.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1714-1714
Author(s):  
William B. Slayton ◽  
Xiao-Miao Li ◽  
Steven M. Guthrie ◽  
Marda L. Jorgensen ◽  
John R. Wingard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Y Chromosome ◽  
Marrow Transplant ◽  
Green Fluorescence ◽  
Hematopoietic Stem ◽  
Post Transplant ◽  
Megakaryocyte Progenitors

Abstract Marrow sinusoidal capillaries provide a niche for megakaryocyte progenitors and possibly hematopoietic stem cells. We sought to determine the fate of host sinusoidal capillaries during marrow transplant. We transplanted whole bone marrow, 2000 sorted Linneg Sca-1pos c-kitpos (SKL) hematopoietic stem and progenitor cells, or single SKL cells (along with rescue marrow) from male mice expressing green fluorescent protein into lethally irradiated female C57/BL6 hosts. We used green fluorescence and the presence of the Y-chromosome to identify donor-derived cells. Sinusoidal engraftment was studied from day 5 up to 1 year post-transplant and in secondary transplants. We identified numerous donor-derived cells based on green fluorescence or presence of the Y chromosome lining the sinusoids, but it was unclear whether these cells were vascular or hematopoietic. Furthermore, we were unable to demonstrate expression of von Willebrand factor, CD31 and MECA-32 using immunohistochemistry in healthy sinusoids. We used two methods to definitively identify donor-derived endothelial cells. First, we identified these cells based on location, characteristic shape and nuclei on H&E stained sections. Serial high power fields were photographed and then the same sections were stained with X and Y FISH probes. Second, we identified donor endothelial cells based on uptake of intravenously injected Ac-LDL which was endocytosed predominantly by sinusoidal endothelial cells four hours after injection. Flushed femoral bone marrow was treated with dispase and collagenase, and disaggregated cells were lineage depleted with a standard lineage cocktail containing Mac-1 antibody. Cytospins were photographed, Ac-LDL staining cells identified, and the same cytospins were stained with X and Y FISH probes. We counted numerous donor-derived endothelial cells whether 2X106 whole bone marrow (n=6 ), 2X104 SKL cells (n=6), or single SKL cells (n=2) were transplanted. These donor-derived endothelial cells were functional based on their ability to uptake DiI AcLDL. These cells appeared adjacent to mature, donor-derived megakaryocytes, suggesting function as a niche for megakaryocyte progenitors. Furthermore, donor-derived endothelial cells were present at levels similar to hematopoietic engraftment in every animal analyzed, suggesting robust levels of repair. Single cell transplants were assessed at seven and nine months, and in secondary transplants, establishing that these cells self-renew. To determine the mechanism whereby HSC’s repair the bone marrow sinusoids, we measured the expression of stromal derived factor-1 (SDF-1) at various time points during the first two weeks post-transplant by semi-quantitative RT-PCR, ELISA, and immunohistochemistry. SDF-1 expression peaked 3 days post-transplant and was expressed primarily by damaged blood vessels. The temporal and spatial pattern of engraftment matched expression patterns of SDF-1. This study demonstrates repair of host sinusoids by donor-derived endothelial cells following transplant, with levels of reconstitution similar to that of the hematopoietic system. This repair may be mediated by expression of SDF-1 by damaged marrow vasculature. Repair of sinusoidal capillaries may be a primary role of the HSC during transplant that is necessary for successful hematopoietic engraftment.

Effect of Umbilical Cord Blood Hematopoietic Stem Cells on Formation of Angiogenic Structures.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4054-4054
Author(s):  
Aaron Victor ◽  
Mary J. Laughlin ◽  
Marcie R. Finney ◽  
Nicholas J. Greco
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Coronary Artery ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Hematopoietic Stem Cell ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Hematopoietic Stem

Abstract There is a significant unmet need for novel therapeutic treatments for patients presenting with chronic ischemic conditions such as coronary artery disease and diabetes. Revascularization measures, such as infusions with endothelial progenitor cells (EPC) characterized by the expression of early hematopoietic stem cell markers, hold significant potential in treating these patients. Pre-clinical and clinical studies using transplanted EPC to restore blood flow and improve cardiac function in animal models of ischemia have proven effective. Recent studies have used bone marrow mononuclear cells while some more recent studies have focused on enriched stem cell treatments, such as purified bone marrow hematopoietic stem cell (HSC) CD34+/133+ cell populations, in patients with coronary artery ischemia. In this study, the hypothesis to be tested was that umbilical cord blood-derived hematopoietic stem cells (CD34+/CD133+) cells may augment the formation and stability of angiogenic networks of cord-like structures derived from umbilical vein endothelial cells (HUVEC) cultured in growth factor-reduced Matrigel (GFR MG) assays. Umbilical cord blood MNC were isolated with ficoll and separated into HSC CD34+/133+ and CD34−/133− fractions. Positive fractions were flow cytometry, sorted for HSC, and stained with the lipophilic fluorescent red dye CM-DiI and the HUVEC were stained with the lipophilic fluorescent green dye Oregon Green. HUVEC alone or HSC and HUVEC were then co-cultured under hypoxic conditions (1% O2) on the GFR MG in 96 well plates. Cells were photographed with a fluorescent microscope at 16, 48, and 72 hours. Transwell experiments (0.4μm pores) were also performed with HSC CD34+/133+ and CD34−/133− fractions prepared and suspended in transwells above HUVEC plated on GFR MG on bottom wells. The presence of both HSC CD34+/133+ and CD34−/133− fractions increased the numbers of nodes (branch points of structures) and allowed the structures to persist when observed over three days (a representative experiment of N =3) (Table): Day 1 Day 1 Day 2 Day 2 Day 3 Day 3 Node # % Total Node # % Total Node # % Total HUVEC 11.6 ± 4.9 100 1.3 ± 1.2 9.2 0.33 ± 0.58 2.2 HUVEC + HSC CD34+/133+ 17.3 ± 9.2 100 6.3 ± 4.5 35.3 4.7 ± 5.5 21.4 HUVEC + HSC CD34−/133− 34 ± 13.2 100 19.7 ± 2.5 61.6 10 ± 3.6 29.8 The HSC CD34−/133− fraction resulted in a greater increase in node formation than the HSC CD34+/133+ and both fractions stimulated significant persistence in formed structures. In addition, CM-Dil labeled cells were localized at nodes points. Results with the transwell assay demonstrated that when either HSC CD34+/133+ or CD34−/133− fractions were suspended above HUVEC, augmentation of the formation of cord-like structures was not observed. In summary, both umbilical cord blood-derived HSC CD34+/133+ and CD34−/133− fractions possess properties that augment the formation of angiogenic structures. We observed that the number of nodes are greater in the presence of both HSC CD34+/133+ and CD34−/133− fractions than with HUVEC alone. The transwell experiment suggested that cell-to-cell interactions are necessary for augmentation of the cord structures. In future studies, we will address the mechanism of intercellular interactions that result in the augmentation of cord-like structures and which particular subpopulations within cord blood, both from HSC CD34+/133+ and CD34−/133− fractions are required for augmentation of structure formation.

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