A simple tool to improve pluripotent stem cell differentiation

Chetty, Sundari; Pagliuca, Felicia W.; Honoré, Christian; Kweudjeu, Anastasie; Rezania, Alireza; Melton, Douglas A.

doi:10.1038/nmeth.2442

Cited by 168 publications

(205 citation statements)

References 36 publications

Supporting

Mentioning

178

Contrasting

Order By: Relevance

“…The yield of cardiomyocytes was, however, quite low, especially with embryoid body-based protocols that patently provided no more that several per cent of robustly beating cells. Different small-molecule approaches to increase the yield were tried and tested, and in 2013 was published a report that treatment of differentiating cells with dimethylsulfoxide (DMSO) dramatically increased the differentiation rates in all germ layer lineages, including the cardiogenic lineage [18]. Fig.…”

Section: Brief Historical Overview Of the Attempts To Cultivate Rhythmentioning

confidence: 99%

“…Treatment with 1-2 % DMSO for 24-28 h was shown to increase the yield of differentiated cells originating from all three germ layers, the cardiogenic lineage included [18]. The authors of the cited study claim that the phenomenon has been observed with > 25 different human embryonic stem cell lines and induced pluripotent stem cell lines.…”

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

“…It has been proposed that the presence of DMSO increases the competence for differentiation in undifferentiated cells by induction of a transient G1 phase arrest [18,149]. Normally, cells of early mammalian embryos and embryonic stem cells cycle rapidly, with significantly shortened G1 phase of the cell cycle.…”

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

“…In late G1, pRB1 is phosphorylated by activated cyclin-dependent kinases in order to release E2F and allow the cell to proceed to S phase of the cell cycle [159,160]. Treatment of differentiating embryonic cells with DMSO resulted in 4-fold increase in the levels of hypophosphorylated retinoblastoma protein [18]. Thus, it is proposed that following DMSO treatment the cell cycle is arrested at the pre-replication G1/S checkpoint, which is a potent trigger of the differentiation programme.…”

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

“…Even among these cell lines that score very high on the 'lineage scorecards' for a particular lineage, the percentage of specialised cells that are produced by targeted differentiation may significantly vary between experiments and between laboratories. Indeed, there have been recent developments in experimental techniques that were shown to increase the differentiation rates for almost all cell types [18], but the consistency of the differentiation rates into the cardiogenic lineage still leaves much to be desired, especially with embryoid bodies-based methodology for differentiation. The latter typically yields no more than 8-10 % of pulsating cardiomyocyte-like cells and about 1-2 % of true cardiomyocytes [13, 200, reviewed in 98].…”

Section: 3mentioning

confidence: 99%

See 4 more Smart Citations

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

Arabadjiev¹,

Petkova²,

Chakarov³

et al. 2014

View full text Add to dashboard Cite

Citation: Arabadjiev A, Petkova R, Chakarov S, Pankov R, Zhelev N. We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells. AbstractHuman cell lines, including disease cell lines are often superior to routine animal models for the purposes of rapid and safe assessment of the effects of different agents that may modulate myocardial functioning under physiological and pathological conditions. There are several currently existing methodologies for derivation of cardiomyocyte-like cells by targeted differentiation from pluripotent cells and by reprogramming/ transdifferentiation from other types of cells (multipotent progenitors, somatic cells, etc). The present paper reviews the potential sources of cells capable of differentiation along the cardiomyocyte lineage; the existing methodologies for targeted differentiation, outlining the specificities of each method; and the markers for differentiation along the mesodermal and the cardiogenic lineage. The yield of robustly beating cells expressing cardio-specific proteins derived by any of the existing methods, however, still rarely exceeds 70-90 %, even with the newly developed approaches for increasing the differentiation capacity. There still is significant variance in the results obtained by different research groups and even between different experiments carried out in the same laboratory, with the same type of cells and same general type of protocol. Derivation of new lines of human pluripotent and multipotent stem cells according to standardised protocols and in completely defined; xeno-free conditions may increase the reliability and reproducibility of research and speed up the development of potential clinical applications.

show abstract

Section: Brief Historical Overview Of the Attempts To Cultivate Rhythmentioning

confidence: 99%

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

Section: Increasing the Propensity For Differentiation Along Multiplementioning

confidence: 99%

Section: 3mentioning

confidence: 99%

See 3 more Smart Citations

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

Arabadjiev¹,

Petkova²,

Chakarov³

et al. 2014

View full text Add to dashboard Cite

show abstract

Scaffolds Functionalized with Matrix from Induced Pluripotent Stem Cell Fibroblasts for Diabetic Wound Healing

Santarella

Sridharan

Marinković

et al. 2020

Adv Healthcare Materials

View full text Add to dashboard Cite

Diabetic foot ulcers (DFUs) are chronic wounds, with 20% of cases resulting in amputation, despite intervention. A recently approved tissue engineering product—a cell‐free collagen‐glycosaminoglycan (GAG) scaffold—demonstrates 50% success, motivating its functionalization with extracellular matrix (ECM). Induced pluripotent stem cell (iPSC) technology reprograms somatic cells into an embryonic‐like state. Recent findings describe how iPSCs‐derived fibroblasts (“post‐iPSF”) are proangiogenic, produce more ECM than their somatic precursors (“pre‐iPSF”), and their ECM has characteristics of foetal ECM (a wound regeneration advantage, as fetuses heal scar‐free). ECM production is 45% higher from post‐iPSF and has favorable components (e.g., Collagen I and III, and fibronectin). Herein, a freeze‐dried scaffold using ECM grown by post‐iPSF cells (Post‐iPSF Coll) is developed and tested vs precursors ECM‐activated scaffolds (Pre‐iPSF Coll). When seeded with healthy or DFU fibroblasts, both ECM‐derived scaffolds have more diverse ECM and more robust immune responses to cues. Post‐iPSF‐Coll had higher GAG, higher cell content, higher Vascular Endothelial Growth Factor (VEGF) in DFUs, and higher Interleukin‐1‐receptor antagonist (IL‐1ra) vs. pre‐iPSF Coll. This work constitutes the first step in exploiting ECM from iPSF for tissue engineering scaffolds.

show abstract

Engineering a Microfluidic Platform to Cryopreserve Stem Cells: A DMSO‐Free Sustainable Approach

Modaresi,

Pacelli,

Chakraborty

et al. 2024

Adv Healthcare Materials

View full text Add to dashboard Cite

Human adipose‐derived stem cells (hASCs) are cryopreserved traditionally using dimethyl sulfoxide (DMSO) as the cryoprotectant agent. DMSO penetrates cell membranes and prevents cellular damage during cryopreservation. However, DMSO is not inert to cells, inducing cytotoxic effects by causing mitochondrial dysfunction, reduced cell proliferation, and impaired hASCs transplantation. Additionally, large‐scale production of DMSO and contamination can adversely impact the environment. A sustainable, green alternative to DMSO is trehalose, a natural disaccharide cryoprotectant agent that does not pose any risk of cytotoxicity. However, the cellular permeability of trehalose is less compared to DMSO. Here, a microfluidic chip is developed for the intracellular delivery of trehalose in hASCs. The chip is designed for mechanoporation, which creates transient pores in cell membranes by mechanical deformation. Mechanoporation allows the sparingly permeable trehalose to be internalized within the cell cytosol. The amount of trehalose delivered intracellularly is quantified and optimized based on cellular compatibility and functionality. Furthermore, whole‐transcriptome sequencing confirms that less than 1% of all target genes display at least a twofold change in expression when cells are passed through the chip compared to untreated cells. Overall, the results confirm the feasibility and effectiveness of using this microfluidic chip for DMSO‐free cryopreservation of hASCs.

show abstract

A simple tool to improve pluripotent stem cell differentiation

Cited by 168 publications

References 36 publications

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

Scaffolds Functionalized with Matrix from Induced Pluripotent Stem Cell Fibroblasts for Diabetic Wound Healing

Engineering a Microfluidic Platform to Cryopreserve Stem Cells: A DMSO‐Free Sustainable Approach

Contact Info

Product

Resources

About