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Cryopreservation of Human Bone Marrow Grafts
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
Evaluation of different freezing methods suffers from the lack of an in vitro assay for the pluripotent hematopoietic stem cells. Culturing of colony-forming progenitor cells after the standard freezing and thawing procedures shows that the survival of these cells is satisfactory.13–17 For example, recovery of CFU-GM has been reported to be ≥71%,14–17 BFU-E (burst-forming-unit-erythroid) ≥72%,15,16 CFU-Mk (colony forming units-megakaryocytes) ≥70%,17 and CFU-GEMM (colony-forming units—granulocyte, erythrocyte, macrophage, and megakaryocyte) ≥89%.17 However, large variations have been observed in the post-thaw survival of progenitor cells of marrow cryopreserved after 4-HC treatment.43 The fact that durable engraftment can be obtained with purged marrow grafts that contained very low numbers of CFU-GM, further emphasizes the inadequacy of the current assay systems.
Cellular Components of Blood
Published in Peter Kam, Ian Power, Michael J. Cousins, Philip J. Siddal, Principles of Physiology for the Anaesthetist, 2020
Peter Kam, Ian Power, Michael J. Cousins, Philip J. Siddal
The PHSCs give rise to all mature blood cells that circulate freely in the peripheral blood. The stem cells undergo cell division and maturation in the bone marrow. The erythroid, granulocytic and megakaryocytic cell lines are derived from a common pluripotential stem cell, appearing as an early myeloid precursor called CFUGEMM (colony-forming unit, granulocyte, erythrocyte, monocyte and megakaryocyte). The precursor cells are stimulated by haemopoietic growth factors, resulting in considerable amplification within the system and increased production of one or more cell lines in accordance to need. The bone marrow is also the primary origin of lymphocytes, and there is some evidence for a common precursor cell for both myeloid and lymphoid cells (Figure 51.1).
Development of an Oligodeoxynucleotide Pharmaceutical for the Treatment of Human Leukemia
Published in Eric Wickstrom, Clinical Trials of Genetic Therapy with Antisense DNA and DNA Vectors, 2020
Alan M. Gewirtz, Deborah Lee Sokol
In this particular study, colony formation was observed in eight of eleven cases evaluated and was statistically significant (p≤0.03) in seven. The amount of inhibition seen was dose dependent and ranged between 58% and 93%. In two cases the effect of the c-myb oligomers on CFU-GEMM colony formation was also determined to assess the effect of the oligomers on progenitors more primitive than CFU-GM. In each case, significant inhibition of CFU-GEMM derived colony formation was noted. Blast cells were isolated from the peripheral blood of AML patients and exposed to sense or antisense oligomers. Colonies and clusters were enumerated and values were compared with growth in control cultures, which contained no oligomer. For each case, the number of colonies or clusters arising in the untreated control dishes was assumed to represent maximal (100%) growth for that patient. The numbers of colonies or clusters arising in the oligomer-treated dishes are expressed as a percentage of this number. NG, no growth. The statistical significance (determined by Student's t test for unpaired samples) of the change observed in the antisense-treated dishes relative to the untreated control is give as a p value in parentheses (reprinted, with permission, from Calabretta et al., 1991).
Enalaprilat and losartan decrease erythroid precursors frequency in cells from patients with polycythemia vera
Published in Hematology, 2023
Angela Bozza, Martina Bernardi, Daniela Catanzaro, Katia Chieregato, Anna Merlo, Giuseppe Astori
PB-MNCs of JAK2V617F positive PV patients treated with enalaprilat showed a significant reduction in BFU-E number at every dose tested. (Figure 3(A)). The number of CFU-GEMM decreased significantly with respect to controls only at 1μg/ml of enalaprilat (Figure 3(A)). Instead, treatment with losartan caused a significant reduction of BFU-E number only at high doses (1 and 10 µg/ml) (Figure 3(B)). CFU-GM did not show any significant change with both drugs, except for 1 µg/ml enalaprilat concentration in which GM number increased (Figure 3(A,B)). We tested also samples isolated from BM of PV patients. In these cells, BFU-E frequency decreased only at the lowest dose of enalaprilat (Figure 3(C)) and 0.1 and 1 µg/ml losartan (Figure 3(D)). No differences were observed in GEMM and GM colonies with both drugs, except for GEMM reduction with 0.01 µg/ml losartan. The reduction in erythroid colony number observed in PB samples correlates with the reduction in total colony number with both drugs, except at 1 µg/ml enalaprilat (Figure 3(E)). In BM samples, this correspondence is not present, probably due to higher variability in GEMM and GM counts (Figure 3(F)).
In vitro expansion of CD 133+ cells derived from umbilical cord blood in poly-L -lactic acid (PLLA) scaffold coated with fibronectin and collagen
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2018
Maryam Islami, Yousef Mortazavi, Masoud Soleimani, Samad Nadri
The 5 × 103 scaffold-expanded cells were harvested from each well before (day 0) and after expansion (day 7). Then, these cells were utilized in 3 ml of methylcellulose medium (H4435, stem cell technology, Canada) plated in a 35-mm culture dish (Grainger). The cell’s dish with two uncovered dishes containing 3–4 ml sterile water was put into a 100-mm culture dish and covered. The sterile water dish was served to steadfastly keep up the humidity essential for colony development. H4435 contains 1% methylcellulose in Iscove's Modified Dulbecco's Medium (IMDM) (IMDM, sigma, Germany). After 14 days incubation at 37°C and 5% CO2, colony-forming unit (CFU)-GM, BFU-E/CFU-E and CFU-GEMM (colony-forming unit of granuiocyte/erythrocyte]macrophage/megakaryocyte; the multilineage HSCs) colonies were counted.
ASXL1 mutations in myeloid neoplasms: pathogenetic considerations, impact on clinical outcomes and survival
Published in Current Medical Research and Opinion, 2018
Juliana Alvarez Argote, Constantin A. Dasanu
Davies et al.27 performed an in vitro study with CD34+ bone marrow cells, and found that silencing ASXL1 expression by lentiviral transduction alters granulomonocytic differentiation. ASXL1 deficiency leads to deficient expression of CD11b and CD15 molecules involved in granulomonocytic differentiation. ASXL1 deficiency in CD34+ cells is also associated with increased number of multipotent mixed lineage colony forming units (CFU-GEMM), granulocyte–macrophage colony forming units (CFU-GM), and granulocyte colony forming units (CFU-G)27. Mouse models showed similar results where the incidence of MDS is increased after ASXL1 gene knockdown28.