Novel Method for Efficient Production of Multipotential Hematopoietic Progenitors from Human Embryonic Stem Cells

2007 ◽  
Vol 85 (5) ◽  
pp. 371-379 ◽  
Author(s):  
Feng Ma ◽  
Dan Wang ◽  
Sachiyo Hanada ◽  
Yasuhiro Ebihara ◽  
Hirohide Kawasaki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Efficient Production ◽  
Hematopoietic Progenitors ◽  
Novel Method

A Novel Method for Efficient Production of Multipotential Hematopoietic Progenitors from Human Embryonic Stem Cells by Co-Culture with Murine Fetal Liver-Derived Stromal Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4214-4214
Author(s):  
Feng Ma ◽  
Dan Wang ◽  
Sachiyo Hanada ◽  
Hirohide Kawasaki ◽  
Yuji Zaike ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Stromal Cells ◽  
Blood Cells ◽  
Fetal Liver ◽  
Hematopoietic Cells ◽  
Embryonic Stem ◽  
Efficient Production ◽  
Hematopoietic Progenitors

Abstract Human embryonic stem cells provide a unique tool to study early events occurring in the development of human embryonic hematopoiesis, and their totipotent capability indicates a potent clinical application based on the cellular therapy and the evaluation of drug effects on hematopoietic and blood cells. To achieve efficient production of hematopoietic cells from human embryonic stem cells, we attempted to reproduce the circumstance surrounding embryonic hematopoietic cells in vitro. Since fetal liver is the predominant source of hematopoietic and blood cells in mammalian embryogenesis, we established stromal cells from mouse fetal liver at days 14 to 15 of gestation. In the co-culture of human embryonic stem cells with the established stromal cells, a number of hematopoietic progenitors were generated at around day 14 of co-culture, and this hematopoietic activity was highly enriched in the cobble stone-like cells under the stromal layer. Most of the cobble stone-like cells collected expressed CD34 and contained a variety of hematopoietic colony-forming cells, especially multilineage colony-forming cells, at a high frequency. The multipotential hematopoietic progenitors in the cobble stone-like cells produced all types of mature blood cells, including adult type hemoglobin-synthesizing erythrocytes and tryptase and chymase-bouble positive mast cells in the suspension cultiue with a cytokine cocktail. The developed co-culture system of human embryonic stem cells should offer a novel source for hematopoietic and blood cells applicable to cellular therapies and drug screening.

Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors

Blood ◽  
2008 ◽  
Vol 111 (11) ◽  
pp. 5298-5306 ◽  
Author(s):  
Naoya Takayama ◽  
Hidekazu Nishikii ◽  
Joichi Usui ◽  
Hiroko Tsukui ◽  
Akira Sawaguchi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Efficient Production ◽  
Hematopoietic Progenitors ◽  
Alternative Source ◽  
Promising Tool ◽  
Spherical Cells

Abstract Human embryonic stem cells (hESCs) could potentially represent an alternative source for blood transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. When we cultured hESCs on either C3H10T1/2 or OP-9 cells to facilitate hematopoiesis, we found that exogenous administration of vascular endothelial growth factor promoted the emergence of sac-like structures, which we named embryonic stem cell–derived sacs (ES-sacs). These ES-sacs consisted of multiple cysts demarcated by cellular monolayers that retained some of the properties of endothelial cells. The spherical cells inside ES-sacs expressed primarily CD34, along with VE-cadherin, CD31, CD41a, and CD45, and were able to form hematopoietic colonies in semisolid culture and to differentiate into mature megakaryocytes by day 24 in the presence of thrombopoietin. Apparently, ES-sacs provide a suitable environment for hematopoietic progenitors. Relatively large numbers of mature megakaryocytes could be induced from the hematopoietic progenitors within ES-sacs, which were then able to release platelets that displayed integrin αIIbβ3 activation and spreading in response to ADP or thrombin. This novel protocol thus provides a means of generating platelets from hESCs, which could serve as the basis for efficient production of platelets for clinical transfusion and studies of thrombopoiesis.

P–797 A novel method for establishing human embryonic stem cells independent of feeder cells

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Cai
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Success Rate ◽  
Cell Lines ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Feeder Cells ◽  
Novel Method

Abstract Study question Is there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells? Summary answer We established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells. What is known already Establishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts) ,this brought in potential heterogeneous pollution.Although there had be some reports about generating human ESCs independent of feeder cells,but the efficiency was low and conditioned medium were unstable and also had the biological contamination. Study design, size, duration We used ten day5/6 donated human blastocysts from our reproductive center ,most of them were genetically diseased embryos with abnormal PGT diagnosis.After establishing ESCs procedure , all the cell lines were identified with pluripotency and differentiation potential tests.The success rate of system was calculated and compared with the conventional methods. Participants/materials, setting, methods In brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells.About 10 days,the typical ES clone can be mechanically passaged and cells can be cultured in normal oxygen concentrations after passage 2. . Main results and the role of chance Using this method we had successfully established nine embryonic stem cell lines from donated human blastocysts ,the success rate was 90% (9/10). Each cell lines had passed the evaluation test of embryonic stem cell. When compared with the conventional feeder cells dependent method,our novol methods not only eliminated the pollution from heterogeneous cells,but also had higher success rate (90% vs 25%). Limitations, reasons for caution Due to the scarcity of donated human blastocysts, this experiment was a single-center experiment with small samples. Wider implications of the findings: We speculated that the batch differences of culture dishes, matrix and culture medium might affect the establish efficiency , and how to carry out a high level of quality control work might be the key factor to keep the system stable. Trial registration number basic research

P-797 A novel method for establishing human embryonic stem cells independent of feeder cells

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Cai
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Success Rate ◽  
Cell Lines ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Feeder Cells ◽  
Novel Method

Abstract Study question Is there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells? Summary answer We established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells. What is known already Establishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts), this brought in potential heterogeneous pollution. Although there had be some reports about generating human ESCs independent of feeder cells, but the efficiency was low and conditioned medium were unstable and also had the biological contamination. Study design, size, duration We used ten day5/6 donated human blastocysts from our reproductive center, most of them were genetically diseased embryos with abnormal PGT diagnosis. After establishing ESCs procedure, all the cell lines were identified with pluripotency and differentiation potential tests. The success rate of system was calculated and compared with the conventional methods. Participants/materials, setting, methods In brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells. About 10 days,the typical ES clone can be mechanically passaged and cells can be cultured in normal oxygen concentrations after passage 2.. Main results and the role of chance Using this method we had successfully established nine embryonic stem cell lines from donated human blastocysts, the success rate was 90% (9/10). Each cell lines had passed the evaluation test of embryonic stem cell. When compared with the conventional feeder cells dependent method,our novol methods not only eliminated the pollution from heterogeneous cells,but also had higher success rate (90% vs 25%). Limitations, reasons for caution Due to the scarcity of donated human blastocysts, this experiment was a single-center experiment with small samples. Wider implications of the findings We speculated that the batch differences of culture dishes, matrix and culture medium might affect the establish efficiency, and how to carry out a high level of quality control work might be the key factor to keep the system stable. Trial registration number basic research

A Feeder-Free and Efficient Production of Functional Neutrophils from Human Embryonic Stem Cells

Stem Cells ◽  
2009 ◽  
Vol 27 (1) ◽  
pp. 59-67 ◽  
Author(s):  
Koichi Saeki ◽  
Kumiko Saeki ◽  
Masako Nakahara ◽  
Satoko Matsuyama ◽  
Naoko Nakamura ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Efficient Production
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