Combinatorial ECM Arrays Identify Cooperative Roles for Matricellular Proteins in Enhancing the Generation of TH+ Neurons From Human Pluripotent Cells

The development of efficient cell culture strategies for the generation of dopaminergic neurons is an important goal for transplantation-based approaches to treat Parkinson’s disease. To identify extracellular matrix molecules that enhance differentiation and might be used in these cell cultures we have used micro-contact printed arrays on glass slides presenting 190 combinations of 19 extracellular matrix molecules selected on the basis of their expression during embryonic development of the ventral midbrain. Using long-term neuroepithelial stem cells (Lt-NES), this approach identified a number of matricellular proteins that enhanced differentiation, with the combination of Sparc, Sparc-like (Sparc-l1) and Nell2 increasing the number of tyrosine hydroxylase+ neurons derived from Lt-NES cells and, critically for further translation, human pluripotent stem cells.

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Correction to: Development of a simple, repeatable, and cost-effective extracellular matrix for long-term xeno-free and feeder-free self-renewal of human pluripotent stem cells

Histochemistry and Cell Biology ◽

10.1007/s00418-021-01963-4 ◽

2021 ◽

Author(s):

Mohammad Pakzad ◽

Mohammad Kazemi Ashtiani ◽

Seyed Latif Mousavi-Gargari ◽

Hossein Baharvand

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Pluripotent Stem Cells ◽

Cost Effective ◽

Human Pluripotent Stem Cells ◽

Self Renewal

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Development of a simple, repeatable, and cost-effective extracellular matrix for long-term xeno-free and feeder-free self-renewal of human pluripotent stem cells

Histochemistry and Cell Biology ◽

10.1007/s00418-013-1144-3 ◽

2013 ◽

Vol 140 (6) ◽

pp. 635-648 ◽

Cited By ~ 8

Author(s):

Mohammad Pakzad ◽

Mohammad Kazemi Ashtiani ◽

Seyed Latif Mousavi-Gargari ◽

Hossein Baharvand

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Pluripotent Stem Cells ◽

Cost Effective ◽

Human Pluripotent Stem Cells ◽

Self Renewal

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Human myocardial extracellular matrix directs the differentiation of pluripotent stem cells toward a cardiomyocyte phenotype

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0034-1367264 ◽

2014 ◽

Vol 62 (S 01) ◽

Author(s):

B. Oberwallner ◽

A. Brodarac ◽

P. Anic ◽

T. Saric ◽

K. Bieback ◽

...

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Pluripotent Stem Cells

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Stability of Imprinting and Differentiation Capacity in Naïve Human Cells Induced by Chemical Inhibition of CDK8 and CDK19

Cells ◽

10.3390/cells10040876 ◽

2021 ◽

Vol 10 (4) ◽

pp. 876

Author(s):

Raquel Bernad ◽

Cian J. Lynch ◽

Rocio G. Urdinguio ◽

Camille Stephan-Otto Attolini ◽

Mario F. Fraga ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Primordial Germ Cell ◽

Normal Karyotype ◽

Cell Types ◽

Dna Demethylation ◽

Starting State ◽

Teratoma Formation

Pluripotent stem cells can be stabilized in vitro at different developmental states by the use of specific chemicals and soluble factors. The naïve and primed states are the best characterized pluripotency states. Naïve pluripotent stem cells (PSCs) correspond to the early pre-implantation blastocyst and, in mice, constitute the optimal starting state for subsequent developmental applications. However, the stabilization of human naïve PSCs remains challenging because, after short-term culture, most current methods result in karyotypic abnormalities, aberrant DNA methylation patterns, loss of imprinting and severely compromised developmental potency. We have recently developed a novel method to induce and stabilize naïve human PSCs that consists in the simple addition of a chemical inhibitor for the closely related CDK8 and CDK19 kinases (CDK8/19i). Long-term cultured CDK8/19i-naïve human PSCs preserve their normal karyotype and do not show widespread DNA demethylation. Here, we investigate the long-term stability of allele-specific methylation at imprinted loci and the differentiation potency of CDK8/19i-naïve human PSCs. We report that long-term cultured CDK8/19i-naïve human PSCs retain the imprinting profile of their parental primed cells, and imprints are further retained upon differentiation in the context of teratoma formation. We have also tested the capacity of long-term cultured CDK8/19i-naïve human PSCs to differentiate into primordial germ cell (PGC)-like cells (PGCLCs) and trophoblast stem cells (TSCs), two cell types that are accessible from the naïve state. Interestingly, long-term cultured CDK8/19i-naïve human PSCs differentiated into PGCLCs with a similar efficiency to their primed counterparts. Also, long-term cultured CDK8/19i-naïve human PSCs were able to differentiate into TSCs, a transition that was not possible for primed PSCs. We conclude that inhibition of CDK8/19 stabilizes human PSCs in a functional naïve state that preserves imprinting and potency over long-term culture.

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Derivation and Long-Term Culture of Transgene-Free Human Induced Pluripotent Stem Cells on Synthetic Substrates

Stem Cells Translational Medicine ◽

10.5966/sctm.2014-0087 ◽

2014 ◽

Vol 3 (12) ◽

pp. 1410-1417 ◽

Cited By ~ 11

Author(s):

Luis Gerardo Villa-Diaz ◽

Jin Koo Kim ◽

Joerg Lahann ◽

Paul H. Krebsbach

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Long Term Culture ◽

Induced Pluripotent

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Chemically defined and xeno-free culture condition for human extended pluripotent stem cells

Nature Communications ◽

10.1038/s41467-021-23320-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Bei Liu ◽

Shi Chen ◽

Yaxing Xu ◽

Yulin Lyu ◽

Jinlin Wang ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Culture Condition ◽

Human Fibroblast ◽

Culture System ◽

Disease Modeling ◽

Directed Differentiation

AbstractExtended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.

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Development of Multilayer Mesenchymal Stem Cell Cell Sheets

International Journal of Translational Medicine ◽

10.3390/ijtm1010002 ◽

2021 ◽

Vol 1 (1) ◽

pp. 4-24

Author(s):

Jun Ochiai ◽

Yutaka Niihara ◽

Joan Oliva

Keyword(s):

Stem Cells ◽

Stromal Cells ◽

Pluripotent Stem Cells ◽

Cell Sheets ◽

Gene Therapies ◽

The Past ◽

Long Term Effect ◽

Induced Pluripotent ◽

New Guidelines

Cell and gene therapies have been developing dramatically over the past decade. To face and adapt to the development of these new therapies, the Food and Drug Administration (FDA) wrote and updated new guidelines from 2016 and keep updating them. Mesenchymal stem cells (MSCs) are the most used cells for treatment, far ahead from the induced pluripotent stem cells (iPSCs), based on registered clinical trials at clinicaltrials.gov. They are widely used because of their differentiation capacity and their anti-inflammatory properties, but some controversies still require clear answers. Additional studies are needed to determine the dosage, the number, and the route of injections (location and transplantation method), and if allogenic MSCs are safe compared to autologous MSC injection, including their long-term effect. In this review, we summarize the research our company is conducting with the adipose stromal cells in engineering cell sheets and their potential application.

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