scholarly journals Who Owns Hematopoietic Stem Cells? Some Fundamental Legal Stakes for the Manufacture of Cultured Red Blood Cells

2017 ◽  
Vol 1 (1) ◽  
Author(s):  
Ythier Jean Mercier
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Hematopoietic Stem

scholarly journals Merocyanine 540-sensitized photoinactivation of human erythrocytes parasitized by Plasmodium falciparum

Blood ◽  
1992 ◽  
Vol 80 (1) ◽  
pp. 21-24 ◽  
Author(s):  
OM Smith ◽  
SA Dolan ◽  
JA Dvorak ◽  
TE Wellems ◽  
F Sieber
Keyword(s):  
Stem Cells ◽  
Plasmodium Falciparum ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Human Blood ◽  
Blood Cells ◽  
Potential Usefulness ◽  
Hematopoietic Stem ◽  
Infected Blood ◽  
Merocyanine 540

The purpose of this study was to evaluate the photosensitizing dye merocyanine 540 (MC540) as a means for extracorporeal purging of Plasmodium falciparum-infected erythrocytes from human blood. Parasitized red blood cells bound more dye than nonparasitized cells, and exposure to MC540 and light under conditions that are relatively well tolerated by normal erythrocytes and normal pluripotent hematopoietic stem cells reduced the concentration of parasitized cells by as much as 1,000-fold. Cells parasitized by the chloroquine- sensitive HB3 clone and the chloroquine-resistant Dd2 clone of P falciparum were equally susceptible to MC540-sensitized photolysis. These data suggest the potential usefulness of MC540 in the purging of P falciparum-infected blood.

Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells

2005 ◽  
Vol 23 (1) ◽  
pp. 69-74 ◽  
Author(s):  
Marie-Catherine Giarratana ◽  
Ladan Kobari ◽  
Hélène Lapillonne ◽  
David Chalmers ◽  
Laurent Kiger ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Ex Vivo ◽  
Hematopoietic Stem ◽  
Human Red Blood Cells

scholarly journals In VitroLarge Scale Production of Human Mature Red Blood Cells from Hematopoietic Stem Cells by Coculturing with Human Fetal Liver Stromal Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
Jiafei Xi ◽  
Yanhua Li ◽  
Ruoyong Wang ◽  
Yunfang Wang ◽  
Xue Nan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Red Blood Cells ◽  
Stromal Cells ◽  
Blood Cells ◽  
Fetal Liver ◽  
Erythroid Differentiation ◽  
Erythroid Cells ◽  
Hematopoietic Stem

In vitromodels of human erythropoiesis are useful in studying the mechanisms of erythroid differentiation in normal and pathological conditions. Here we describe an erythroid liquid culture system starting from cord blood derived hematopoietic stem cells (HSCs). HSCs were cultured for more than 50 days in erythroid differentiation conditions and resulted in a more than 109-fold expansion within 50 days under optimal conditions. Homogeneous erythroid cells were characterized by cell morphology, flow cytometry, and hematopoietic colony assays. Furthermore, terminal erythroid maturation was improved by cosculturing with human fetal liver stromal cells. Cocultured erythroid cells underwent multiple maturation events, including decrease in size, increase in glycophorin A expression, and nuclear condensation. This process resulted in extrusion of the pycnotic nuclei in up to 80% of the cells. Importantly, they possessed the capacity to express the adult definitiveβ-globin chain upon further maturation. We also show that the oxygen equilibrium curves of the cord blood-differentiated red blood cells (RBCs) are comparable to normal RBCs. The large number and purity of erythroid cells and RBCs produced from cord blood make this method useful for fundamental research in erythroid development, and they also provide a basis for future production of available RBCs for transfusion.

scholarly journals Merocyanine 540-sensitized photoinactivation of human erythrocytes parasitized by Plasmodium falciparum

Blood ◽  
1992 ◽  
Vol 80 (1) ◽  
pp. 21-24 ◽  
Author(s):  
OM Smith ◽  
SA Dolan ◽  
JA Dvorak ◽  
TE Wellems ◽  
F Sieber
Keyword(s):  
Stem Cells ◽  
Plasmodium Falciparum ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Human Blood ◽  
Blood Cells ◽  
Potential Usefulness ◽  
Hematopoietic Stem ◽  
Infected Blood ◽  
Merocyanine 540

Abstract The purpose of this study was to evaluate the photosensitizing dye merocyanine 540 (MC540) as a means for extracorporeal purging of Plasmodium falciparum-infected erythrocytes from human blood. Parasitized red blood cells bound more dye than nonparasitized cells, and exposure to MC540 and light under conditions that are relatively well tolerated by normal erythrocytes and normal pluripotent hematopoietic stem cells reduced the concentration of parasitized cells by as much as 1,000-fold. Cells parasitized by the chloroquine- sensitive HB3 clone and the chloroquine-resistant Dd2 clone of P falciparum were equally susceptible to MC540-sensitized photolysis. These data suggest the potential usefulness of MC540 in the purging of P falciparum-infected blood.

Ultra-High-Yield Manufacture of Red Blood Cells from Hematopoietic Stem Cells

2011 ◽  
Vol 17 (11) ◽  
pp. 1131-1137 ◽  
Author(s):  
Nicholas E. Timmins ◽  
Stavrosia Athanasas ◽  
Marko Günther ◽  
Penelope Buntine ◽  
Lars K. Nielsen
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Red Blood Cells ◽  
Blood Cells ◽  
High Yield ◽  
Hematopoietic Stem

scholarly journals Rapid and sustained hematopoietic recovery in lethally irradiated mice transplanted with purified Thy-1.1lo Lin-Sca-1+ hematopoietic stem cells

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3758-3779 ◽  
Author(s):  
N Uchida ◽  
HL Aguila ◽  
WH Fleming ◽  
L Jerabek ◽  
IL Weissman
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Blood Cells ◽  
Critical Role ◽  
White Blood Cells ◽  
Cell Types ◽  
Dose Dependence ◽  
Hematopoietic Stem ◽  
Hematopoietic Recovery

Abstract Hematopoietic stem cells (HSCs) are believed to play a critical role in the sustained repopulation of all blood cells after bone marrow transplantation (BMT). However, understanding the role of HSCs versus other hematopoietic cells in the quantitative reconstitution of various blood cell types has awaited methods to isolate HSCs. A candidate population of mouse HSCs, Thy-1.1lo Lin-Sca-1+ cells, was isolated several years ago and, recently, this population has been shown to be the only population of BM cells that contains HSCs in C57BL/Ka-Thy-1.1 mice. As few as 100 of these cells can radioprotect 95% to 100% of irradiated mice, resulting long-term multilineage reconstitution. In this study, we examined the reconstitution potential of irradiated mice transplanted with purified Thy-1.1lo Lin-Sca-1+ BM cells. Donor-derived peripheral blood (PB) white blood cells were detected as early as day 9 or 10 when 100 to 1,000 Thy-1.1lo Lin-Sca-1+ cells were used, with minor dose-dependent differences. The reappearance of platelets by day 14 and thereafter was also seen at all HSC doses (100 to 1,000 cells), with a slight dose-dependence. All studied HSC doses also allowed RBC levels to recover, although at the 100 cell dose a delay in hematocrit recovery was observed at day 14. When irradiated mice were transplanted with 500 Thy-1.1lo Lin-Sca-1+ cells compared with 1 x 10(6) BM cells (the equivalent amount of cells that contain 500 Thy-1.1lo Lin-Sca-1+ cells as well as progenitor and mature cells), very little difference in the kinetics of recovery of PB, white blood cells, platelets, and hematocrit was observed. Surprisingly, even when 200 Thy1.1lo Lin-Sca- 1+ cells were mixed with 4 x 10(5) Sca-1- BM cells in a competitive repopulation assay, most of the early (days 11 and 14) PB myeloid cells were derived from the HSC genotype, indicating the superiority of the Thy-1.1lo Lin-Sca-1+ cells over Sca-1- cells even in the early phases of myeloid reconstitution. Within the Thy-1.1lo Lin-Sca-1+ population, the Rhodamine 123 (Rh123)hi subset dominates in PB myeloid reconstitution at 10 to 14 days, only to be overtaken by the Rh123lo subset at 3 weeks and thereafter. These findings indicate that HSCs can account for the early phase of hematopoietic recovery, as well as sustained hematopoiesis, and raise questions about the role of non-HSC BM populations in the setting of BMT.

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.

Resistance of Hematopoietic Stem Cells Lacking the KIR Ligand to Autologous NK Cell Attacks in Patients with Acquired Aplastic Anemia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2939-2939
Author(s):  
Hiroyuki Maruyama ◽  
Luis J. Espinoza ◽  
Takamasa Katagiri ◽  
Yoshitaka Zaimoku ◽  
Koichi Kashiwase ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Aplastic Anemia ◽  
Nk Cells ◽  
Blood Cells ◽  
Nk Cell ◽  
Gene Repertoire ◽  
Malignant Cells ◽  
Hematopoietic Stem ◽  
Accessory Molecules

Abstract Normal blood cells, including hematopoietic stem cells (HSCs), express KIR ligands (KIR-Ls) to protect themselves from an autologous NK cell attack, and malignant cells lacking KIR-Ls elicit NK cell-mediated killing of themselves. This missing-self mechanism is believed to play an important role in the elimination of malignant cells. However, the mechanisms underlying the killing of KIR-L-lacking malignant cells by NK cells remain unclear due to the heterogeneity of tumor cells in terms of their proliferative capacity, and also because other accessory molecules may be involved in the NK cell attacks, in addition to KIR-Ls. This makes it difficult to clarify the interaction between NK cells and KIR-L-lacking target cells. The lack of class I HLA occurs not only in malignant blood cells, but also in the normal leukocytes of patients with acquired aplastic anemia (AA). These HLA-lacking leukocytes, detectable in 13% of patients with AA, are derived from HSCs that undergo copy number neutral loss of heterozygosity of the short arm of chromosome 6 (6pLOH), and thereby escape the cytotoxic T-cell (CTL) attack against HSCs. The 6pLOH may involve KIR-L loss in some patients, leading to a change in the susceptibility of the affected HSCs to NK cell-mediated killing. Unlike malignant cells, HLA-lacking leukocytes are essentially the same as the wild-type leukocytes, except for the HLA expression. Studying 6pLOH (+) AA patients with leukocytes lacking KIR-Ls should therefore be useful for clarifying the roles of KIR-Ls and other accessory molecules in the target cell killing by NK cells. Screening of 389 patients with AA using flow cytometry and a SNP array analysis revealed that there were HLA-A allele-lacking leukocytes in 60 (15.4%) patients, which included 36 C1/C2 and 24 Bw4/Bw6 heterozygotes. Unexpectedly, a lack of KIR-Ls as a result of 6pUPD was found in five patients (13.9%, C1 missing in two and C2 missing in three) of the 36 C1/C2 heterozygotes and in five (20.8%) of the 24 Bw4/Bw6 heterozygotes, although the proportion of patients lacking a KIR-L-containing haplotype (20.8%) was significantly lower than that of patients lacking a haplotype that did not contain KIR-Ls (79.2%). Moreover, the median percentage of HLA-A-lacking granulocytes in the 10 patients who lacked a KIR-L-containing haplotype (12.4%, 0.44%-50.3%) was significantly lower than that (55.3%, 1.4%-99.4%) in the 26 patients who lacked a haplotype that did not contain KIR-Ls, suggesting that the HSCs lacking KIR-Ls or their progenies are susceptible to autologous NK cells to some extent, but are not eliminated completely. To clarify the mechanisms underlying the HSC resistance to NK cells, we determined the KIR gene repertoire and the haplotype of seven patients whose 6pLOH(+) leukocytes were lacking a KIR-L-containing haplotype. All patients possessed inhibitory KIR genes responsive to corresponding KIR-Ls, a finding that negates the possibility that NK cells failed to undergo licensing in these patients. Although the frequency of the KIR-B haplotype, a haplotype associated with a higher cytotoxic function of NK cells, in the seven patients was lower (14%) than that in Japanese healthy individuals (40.1%), two patients possessed the KIR-B haplotype. Phenotypic analyses of the NK cell subsets defined by anti-2DL1, anti-2DL2/2DL3 and anti-3DL1 antibodies showed that all seven patients had a similar percentage of the eight different NK cell subsets, which included 0.5 to 8% of effector NK cells capable of killing leukocytes lacking corresponding KIR-Ls. The expression level of HLA-E was comparable between HLA-A-lacking and HLA-A-retaining monocytes. The expression of NKG2A on the effector NK cells was also comparable to that of the other NK cell subsets in the 6pLOH(+) patients. Our study demonstrated, for the first time, that HSCs lacking KIR-Ls can evade autologous NK cell attack through an as yet unknown mechanism(s) and can continue to generate blood cells in patients with AA. Disclosures No relevant conflicts of interest to declare.

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