Cloning from stem cells: different lineages, different species, same story

2009 ◽  
Vol 21 (1) ◽  
pp. 83 ◽  
Author(s):  
Björn Oback
Keyword(s):  
Stem Cells ◽  
Recipient Cell ◽  
Donor Cell ◽  
Limiting Factor ◽  
Cloning Efficiency ◽  
Surrogate Mothers ◽  
Divergent Differentiation ◽  
Mouse Cells ◽  
Differentiation Status ◽  
Donor Cell Type

Following nuclear transfer (NT), the most stringent measure of extensive donor cell reprogramming is development into viable offspring. This is referred to as cloning efficiency and quantified as the proportion of cloned embryos transferred into surrogate mothers that survive into adulthood. Cloning efficiency depends on the ability of the enucleated recipient cell to carry out the reprogramming reactions (‘reprogramming ability’) and the ability of the nuclear donor cell to be reprogrammed (‘reprogrammability’). It has been postulated that reprogrammability of the somatic donor cell epigenome is inversely proportional to its differentiation status. In order to test this hypothesis, reprogrammability was compared between undifferentiated stem cells and their differentiated isogenic progeny. In the mouse, cells of divergent differentiation status from the neuronal, haematopoietic and skin epithelial lineage were tested. In cattle and deer, skeletal muscle and antler cells, respectively, were used as donors. No conclusive correlation between differentiation status and cloning efficiency was found, indicating that somatic donor cell type may not be the limiting factor for cloning success. This may reflect technical limitations of the NT-induced reprogramming assay. Alternatively, differentiation status and reprogrammability may be unrelated, making all cells equally difficult to reprogramme once they have left the ground state of pluripotency.

Abstract P404: Membrane Composition Of Cardiac-derived Extracellular Vesicles Changes With Vesicle Origin And Determines Uptake

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Sruti Bheri ◽  
Jessica R Hoffman ◽  
Hyun-Ji Park ◽  
Michael E Davis
Keyword(s):  
Extracellular Vesicles ◽  
Lipid Membrane ◽  
Recipient Cell ◽  
Donor Cell ◽  
Membrane Lipid ◽  
Least Squares Regression ◽  
Membrane Composition ◽  
Cell Type ◽  
Uptake Mechanism ◽  
Donor Cell Type

Introduction: Myocardial infarction (MI) is a leading cause of mortality worldwide. The potency of cell-based therapies for MI is increasingly attributed to the release of extracellular vesicles (EVs) which consist of a lipid/protein membrane and encapsulate RNA cargo. Specifically, EVs from ckit+ progenitor cells (CPCs) and mesenchymal stromal cells (MSCs) are shown to be pro-reparative, with clinical trials ongoing. Despite copious research into EV cargo, the role of donor cell type on EV membrane composition and its effects on EV uptake mechanism by recipient cells remain unclear. This is crucial for designing EV-based therapeutics as uptake mechanism dictates the functionality of the cargo. Thus, we hypothesized that (1) EV membrane composition varies by donor cell type and (2) this variation covaries with the mechanism of uptake. Methods: EVs were isolated using differential ultracentrifugation from four cardiac cell types: CPCs, MSCs, cardiac endothelial cells (CECs) and rat cardiac fibroblasts (RCFs) grown in normoxia (18% O 2 ) or hypoxia (1% O 2 ) to mimic ischemic conditions. EVs were characterized for size and concentration. EV lipid membrane profile was assessed through LC/MS/MS. Donor cell’s role on EV uptake mechanism was determined by inhibiting known uptake pathways (clathrin, dynamin, macropinocytosis and caveolae/lipid raft) with small molecules and quantifying CEC/RCF endocytosis of EVs with flow cytometry. Finally, partial least squares regression was used to determine the most important lipids involved in EV uptake mechanism. Results: EVs were successfully isolated and characterized. The EV membrane lipid profiles clustered by donor cell type. Uptake mechanism of EVs varied based on both donor and recipient cell type with dynamin mediated endocytosis being the most common. Further, the uptake mechanism was independent of normoxic/hypoxic conditioning. Finally, supervised learning methods revealed specific lipid classes (sphingolipids and glycerophospholipids) covaried with EV uptake mechanism. Conclusion: This work highlights the importance of the understudied EV membrane and its role in delivering therapeutic cargo. Active donor cell selection for efficient EV uptake will allow for more potent EV-based MI therapies.

Red blood cell generation from human induced pluripotent stem cells of different donor cell type origin

2013 ◽  
Vol 41 (8) ◽  
pp. S27
Author(s):  
Isabel Dorn ◽  
Katharina Klich ◽  
Martina Radstaak ◽  
Katherina Psathaki ◽  
Marcos Arauzo-Bravo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Blood Cell ◽  
Induced Pluripotent Stem Cells ◽  
Red Blood Cell ◽  
Pluripotent Stem Cells ◽  
Donor Cell ◽  
Cell Generation ◽  
Cell Type ◽  
Induced Pluripotent ◽  
Donor Cell Type

Coordination between donor cell type and cell cycle stage improves nuclear cloning efficiency in cattle

2003 ◽  
Vol 59 (1) ◽  
pp. 45-59 ◽  
Author(s):  
D.N Wells ◽  
G Laible ◽  
F.C Tucker ◽  
A.L Miller ◽  
J.E Oliver ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Donor Cell ◽  
Cloning Efficiency ◽  
Cell Type ◽  
Cell Cycle Stage ◽  
Donor Cell Type

scholarly journals Erythroid differentiation of human induced pluripotent stem cells is independent of donor cell type of origin

Haematologica ◽  
2014 ◽  
Vol 100 (1) ◽  
pp. 32-41 ◽  
Author(s):  
I. Dorn ◽  
K. Klich ◽  
M. J. Arauzo-Bravo ◽  
M. Radstaak ◽  
S. Santourlidis ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Donor Cell ◽  
Erythroid Differentiation ◽  
Cell Type ◽  
Induced Pluripotent ◽  
Donor Cell Type

scholarly journals Donor cell type can influence the epigenome and differentiation potential of human induced pluripotent stem cells

2011 ◽  
Vol 29 (12) ◽  
pp. 1117-1119 ◽  
Author(s):  
Kitai Kim ◽  
Rui Zhao ◽  
Akiko Doi ◽  
Kitwa Ng ◽  
Juli Unternaehrer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Donor Cell ◽  
Cell Type ◽  
Differentiation Potential ◽  
Induced Pluripotent ◽  
Donor Cell Type

scholarly journals Dynamic integration of enteric neural stem cells in ex vivo organotypic colon cultures

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Georgina Navoly ◽  
Conor J. McCann
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
Organotypic Culture ◽  
Ex Vivo ◽  
Donor Cell ◽  
Colonic Tissue ◽  
Cell Therapies ◽  
Enteric Ganglia ◽  
Donor Cells

AbstractEnteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remain unclear. Therefore, we aimed to examine ENSC integration using an adapted ex vivo organotypic culture system. Donor ENSC were obtained from Wnt1cre/+;R26RYFP/YFP mice allowing specific labelling, selection and fate-mapping of cells. YFP+ neurospheres were transplanted to C57BL6/J (6–8-week-old) colonic tissue and maintained in organotypic culture for up to 21 days. We analysed and quantified donor cell integration within recipient tissues at 7, 14 and 21 days, along with assessing the structural and molecular consequences of ENSC integration. We found that organotypically cultured tissues were well preserved up to 21-days in ex vivo culture, which allowed for assessment of donor cell integration after transplantation. Donor ENSC-derived cells integrated across the colonic wall in a dynamic fashion, across a three-week period. Following transplantation, donor cells displayed two integrative patterns; longitudinal migration and medial invasion which allowed donor cells to populate colonic tissue. Moreover, significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia. These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease.

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