Developing a co-culture system for effective megakaryo/thrombopoiesis from umbilical cord blood hematopoietic stem/progenitor cells

Hatami, Javad; Andrade, Pedro de; Matos, A. P. Alves de; Djokovic, Dusan; Lilaia, Carla; Ferreira, Frederico Castelo; Silva, Cláudia L. da

doi:10.1016/j.jcyt.2014.12.010

Cited by 10 publications

(3 citation statements)

References 56 publications

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“…A primary thrust of investigations focused on megakaryopoiesis and thrombopoiesis has been to identify culture mediums, cytokine cocktails, and extracellular matrices that yield high numbers of platelets with morphologic characteristics and functions similar to platelets that are freshly isolated from the bloodstream. Results from these studies indicate that slight alterations in the in vitro culture conditions and/or constituents can significantly influence the generation and phenotype of newly formed platelets [17][18][19][20][21][22][23]. This strongly suggests that changes in the bone marrow milieu can alter transcriptional, translational, and post-translational processes throughout megakaryocyte development, differentiation, and proplatelet formation.…”

Section: Megakaryopoiesis and Thrombopoiesis: Key Determinants Of Thementioning

confidence: 96%

Regulation of the genetic code in megakaryocytes and platelets

Rondina

Weyrich

2015

Journal of Thrombosis and Haemostasis

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Summary Platelets are generated from nucleated precursors referred to as megakaryocytes. The formation of platelets is one of the most elegant and unique developmental processes in eukaryotes. Because they enter the circulation without nuclei, platelets are often considered simple, non-complex cells that have limited functions beyond halting blood flow. However, emerging evidence over the past decade demonstrates that platelets are more sophisticated than previously considered. Platelets carry a rich repertoire of messenger RNAs (mRNAs), microRNAs (miRNAs), and proteins that contribute to primary (adhesion, aggregation, secretion) and alternative (immune regulation, RNA transfer, translation) functions. It is also becoming increasing clear that the “genetic code” of platelets changes with race, genetic disorders, or disease. Changes in the “genetic code” can occur at multiple points including megakaryocyte development, platelet formation, or in circulating platelets. This review focuses on regulation of the “genetic code” in megakaryocytes and platelets and its potential contribution to health and disease.

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Section: Megakaryopoiesis and Thrombopoiesis: Key Determinants Of Thementioning

confidence: 96%

Regulation of the genetic code in megakaryocytes and platelets

Rondina

Weyrich

2015

Journal of Thrombosis and Haemostasis

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“…BM mononuclear cells were isolated from BM using lymphocyte separation medium (HaoYang, Tianjin, China) and were seeded into 6-well plates at a concentration of up to 5 £ 10 6 cells per well in DMEM with 10% FBS (Gibco, Grand Island, NY) [24,32,35]. Nonadherent cells were removed after 4 days.…”

Section: Bm Mscs Culturementioning

confidence: 99%

Dysfunctional Bone Marrow Mesenchymal Stem Cells in Patients with Poor Graft Function after Allogeneic Hematopoietic Stem Cell Transplantation

Song

Zhao

Lyu

et al. 2018

Biology of Blood and Marrow Transplantation

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Poor graft function (PGF) is a life-threatening complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and is characterized by defective hematopoiesis. Mesenchymal stem cells (MSCs) have been shown to support hematopoiesis, but little is known about the role of MSCs in the pathogenesis of PGF. In the current prospective case-control study, we evaluated whether the number and function of bone marrow (BM) MSCs in PGF patients differed from those in good graft function (GGF) patients. We found that BM MSCs from PGF patients expanded more slowly and appeared flattened and larger, exhibiting more apoptosis and senescence than MSCs from GGF patients. Furthermore, increased intracellular reactive oxygen species, p-p53, and p21 (but not p38) levels were detected in MSCs from PGF patients. Moreover, the ability of MSCs to sustain hematopoiesis was significantly reduced in PGF patients, as evaluated by cell number, apoptosis, and the colony-forming unit-plating efficiency of CD34 cells. In summary, the biologic characteristics of PGF MSCs are different from those of GGF MSCs, and the in vitro hematopoiesis-supporting ability of PGF MSCs is significantly lower. Although requiring further validation, our study indicates that reduced and dysfunctional BM MSCs may contribute to deficient hematopoiesis in PGF patients. Therefore, improvement of BM MSCs may represent a promising therapeutic approach for PGF patients after allo-HSCT.

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“…[13][14][15][16] Various studies have used MSCs as a feeder layer for the ex vivo expansion and differentiation of HSCs toward megakaryocytic progenitors. 14,[17][18][19] MSCs produce low levels of thrombopoietin (TPO), which synergizes with other cytokines such as IL-6, IL-11, and stem cell factor (SCF) for megakaryocyte differentiation in the absence of exogenous cytokines. 20,21 Furthermore, MSCs secrete different types of membranous particles including exosomes, microvesicles (MVs), and apoptotic bodies into extracellular space.…”

Section: Introductionmentioning

confidence: 99%

Effect of Mesenchymal Stem Cell-derived Microvesicles on Megakaryocytic Differentiation of CD34+ Hematopoietic Stem Cells

et al. 2020

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Purpose: Mesenchymal stem cells (MSCs) release hematopoietic cytokines, growth factors, and Microvesicles (MVs) supporting the hematopoietic stem cells (HSCs). MVs released from various cells, playing a crucial role in biological functions of their parental cells. MSC-derived MVs contain microRNAs and proteins with key roles in the regulation of hematopoiesis. Umbilical cord blood (UCB) is a source for transplantation but the long-term recovery of platelets is a main problem. Therefore, we intend to show that MSC-MVs are able to improve the differentiation of UCB-derived CD34+ cells to megakaryocyte lineage. Methods: In this descriptive study, MSCs were cultured in DMEM to collect the culture supernatant, which was ultracentrifuged for the isolation of MVs. HSCs were isolated from UCB using MACS method and cultured in IMDM supplemented with cytokines and MVs in three different conditions. Megakaryocyte differentiation was evaluated through the expression of specific markers and genes after 72 hours, and the data was analyzed by t test (P<0.05). Results: The expression of specific megakaryocyte markers (CD41 and CD61) in the presence of different concentrations of MSC-MVs did not show any significant difference. Also, the expression of specific genes of megakaryocyte lineage was compared with control group. The expression of GATA2 and c-Mpl was significantly increased, GATA1 was not significantly decreased, and FLI1 was significantly decreased. Conclusion: MSC-MVs could improve the expression of specific megakaryocyte genes; however, there was no significant expression of CD markers. Further studies, including the evaluation of late stages of megakaryocyte differentiation, are required to evaluate platelet production and shedding

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Developing a co-culture system for effective megakaryo/thrombopoiesis from umbilical cord blood hematopoietic stem/progenitor cells

Cited by 10 publications

References 56 publications

Regulation of the genetic code in megakaryocytes and platelets

Regulation of the genetic code in megakaryocytes and platelets

Dysfunctional Bone Marrow Mesenchymal Stem Cells in Patients with Poor Graft Function after Allogeneic Hematopoietic Stem Cell Transplantation

Effect of Mesenchymal Stem Cell-derived Microvesicles on Megakaryocytic Differentiation of CD34+ Hematopoietic Stem Cells

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