Identification of Potential Pluripotency Determinants for Human Embryonic Stem Cells Following Proteomic Analysis of Human and Mouse Fibroblast Conditioned Media

2007 ◽  
Vol 6 (9) ◽  
pp. 3796-3807 ◽  
Author(s):  
Andrew B. J. Prowse ◽  
Leon R. McQuade ◽  
Katherine J. Bryant ◽  
Helder Marcal ◽  
Peter P. Gray
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Proteomic Analysis ◽  
Embryonic Stem ◽  
Mouse Fibroblast ◽  
Conditioned Media ◽  
Human And Mouse

High throughput cryopreservation of cells by rapid freezing of sub-μl drops using inkjet printing – cryoprinting

Lab on a Chip ◽  
2015 ◽  
Vol 15 (17) ◽  
pp. 3503-3513 ◽  
Author(s):  
Rui Dou ◽  
Rachel E. Saunders ◽  
Lisa Mohamet ◽  
Christopher M. Ward ◽  
Brian Derby
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Human Embryonic Stem Cells ◽  
Inkjet Printing ◽  
Embryonic Stem ◽  
Mouse Fibroblast ◽  
Rapid Freezing ◽  
Fibroblast Cells

We have successfully used inkjet printing to cryopreserve 3T3 mouse fibroblast cells and human neuroprogenitor cells (NPCs) derived from human embryonic stem cells (hESCs).

Human embryonic stem cells maintained in the absence of mouse embryonic fibroblasts or conditioned media are capable of hematopoietic development

Blood ◽  
2005 ◽  
Vol 105 (12) ◽  
pp. 4598-4603 ◽  
Author(s):  
Lisheng Wang ◽  
Li Li ◽  
Pablo Menendez ◽  
Chantal Cerdan ◽  
Mickie Bhatia
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Conditioned Media ◽  
Proliferative Potential ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Self Renewal ◽  
Hematopoietic Development

Abstract To date, hematopoietic development of human embryonic stem cells (hESCs) has been limited to cell lines cultured in the presence of either mouse embryonic fibroblasts (MEFs) or MEF-conditioned media (MEF-CM). Anonymous xenogenic factors from MEFs or MEF-CM complicate studies of hESC self-renewal and also raise concerns for the potential clinical applications of generating primitive hematopoietic cells from hESC lines maintained under these ambiguous conditions. Here, we demonstrate that hESCs can be cultured over 30 passages in defined conditions in the absence of MEFs or MEF-CM using only serum replacement (SR) media and high concentrations of basic fibroblast growth factor (SR-bFGF). Similar to hESCs cultured in MEF-CM, hESCs cultured in SR-bFGF sustained characteristics of undifferentiated hESCs, proliferative potential, normal karyotype, in vitro and in vivo 3 germ-layer specification and gave rise to hemogenic-endothelial precursors required for subsequent primitive hematopoietic development. Our report demonstrates that anonymous factors produced by feeder cells are not necessary for hESC maintenance and subsequent hematopoietic specification, thereby providing a defined system for studies of hESC self-renewal and hESC-derived hematopoiesis. (Blood. 2005;105:4598-4603)

scholarly journals StemCell Keep™ is Effective for Cryopreservation of Human Embryonic Stem Cells by Vitrification

2017 ◽  
Vol 26 (5) ◽  
pp. 773-787 ◽  
Author(s):  
Akemi Ota ◽  
Kazuaki Matsumura ◽  
Jun-Jae Lee ◽  
Shoichiro Sumi ◽  
Soung-Hyu Hyon
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cell Number ◽  
Mouse Fibroblast ◽  
Stem Cell Markers ◽  
Rt Pcr ◽  
Induced Pluripotent ◽  
Attachment Ability

Safe and stable cryopreservation is critical for research involving human embryonic stem cells (hESCs). Dimethyl sulfoxide (DMSO) is a popular cryoprotective agent; however, its cytotoxicity cannot be ignored. Thus, there is a need for an alternate cryoprotectant. We reported previously that a novel cryopreservation reagent, StemCell Keep™ (SCK), was effective for cryopreserving human induced pluripotent stem cells (hiPSCs) by vitrification. Because hESCs and hiPSCs are not identical, the current study examined the use of SCK on hESCs. hESCs cryopreserved with SCK were thawed and cultured on SNL 76/7 cells, which were derived from a mouse fibroblast STO cell line transformed with neomycin resistance and murine LIF genes. After cryopreservation, cultured hESCs were assessed for their attachment ability and characterized by alkaline phosphatase (AP) and immunocytochemical (ICC) staining, fluorescence-activated cell sorting (FACS), reverse transcription polymerase chain reaction (RT-PCR), and karyotyping. The proliferation of SCK-cryopreserved hESCs cultured on SNL cells, or in feeder-free conditions, was higher than that of cells preserved in a solution of 2 M DMSO, 1 M acetamide, and 3 M propylene glycol (DAP). The cell number with SCK-cryopreserved hESCs was about twice that of hESCs cryopreserved in DAP. The pluripotency of SCK-cryopreserved hESCs was similar to that of DAP-cryopreserved hESCs based on AP staining. Data from ICC, FACS, and RT-PCR analyses showed that stem cell markers were continually expressed on SCK-cryopreserved hESCs. The teratoma assay showed that SCK-cryopreserved hESCs differentiated into three germ layers. Furthermore, SCK-cryopreserved hESCs had normal karyotypes. These data indicate that SCK was effective for cryopreservation of hESCs by vitrification.

scholarly journals A large-scale proteomic analysis of human embryonic stem cells

BMC Genomics ◽  
2007 ◽  
Vol 8 (1) ◽  
pp. 478 ◽  
Author(s):  
Thomas C Schulz ◽  
Anna Swistowska ◽  
Ying Liu ◽  
Andrzej Swistowski ◽  
Gail Palmarini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Proteomic Analysis ◽  
Large Scale ◽  
Embryonic Stem

Maintenance of Human Embryonic Stem Cells in Mesenchymal Stem Cell-Conditioned Media Augments Hematopoietic Specification

2012 ◽  
Vol 21 (9) ◽  
pp. 1549-1558 ◽  
Author(s):  
Verónica Ramos-Mejía ◽  
Agustín F. Fernández ◽  
Verónica Ayllón ◽  
Pedro J. Real ◽  
Clara Bueno ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Mesenchymal Stem Cell ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Conditioned Media

A proteome analysis of conditioned media from human neonatal fibroblasts used in the maintenance of human embryonic stem cells

PROTEOMICS ◽  
2005 ◽  
Vol 5 (4) ◽  
pp. 978-989 ◽  
Author(s):  
Andrew B. J. Prowse ◽  
Leon R. McQuade ◽  
Katherine J. Bryant ◽  
Derek D. Van Dyk ◽  
Bernard E. Tuch ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Proteome Analysis ◽  
Conditioned Media
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