GMP-ready hepatocyte differentiation protocol 

The method presented here by the group of Dr. David Hay, University of Edinburgh, UK, describes a scalable and GMP-ready differentiation system to generate human hepatocyte-like cells from pluripotent stem cells on human recombinant laminin substrates. It serves as a cost-effective and standardized system to generate human hepatocyte-like cells for basic and applied human liver research.


Get the hepatocyte differentiation protocol here

                                                                                                  2017-03-02

GMP differentiation protocol for stem cell derived hepatocytes on laminin

Efficient production of hESC-derived dopaminergic neurons under GMP conditions

Here, Agnete presents a GMP-compliant protocol for neuralising hESCs directly from day 0 of differentiation. This approach not only enables us to get highly pure populations of DA progenitor cells (>90%), but it also increases our final yield of transplantable cells >40 times when compared to previous embryoid body protocols starting with the same number of cells. This means that starting from just a single 6-well plate of undifferentiated hESCs, we are now able to produce cells in a scale suitable for clinical production. The GMP-compliant dopaminergic differentiation protocol is published in Cell Stem Cell. Enjoy!


Dr. Agnete Kirkeby
PostDoc, Lund University, Sweden

                                                                                                  2015-04-30

 

 

Dopaminergic neuron differentiation under GMP conditions on laminin

MESSENGER RNA REPROGRAMMING
- PATH TO CLINICALLY RELEVANT IPSCS

This talk will review the development of mRNA reprogramming from its inception, describe the particular challenges entailed, and address the latest advances which have facilitated implementation of a high-throughput, xeno-free, feeder-free, footprint-free iPSC derivation pipeline based on this exciting technology.


Dr. Luigi Warren
CEO, Cellular Reprogramming, Inc, USA

                                                                                                  2015-04-30

mRNA reprogramming of clinically compliant iPSC on laminin

Efficient hepatocyte specification
- improved maturation and functional organization

Here, Kate presents her work on that show that culture of human ES cells on human recombinant laminin-521 and laminin-111 substrates significantly improve hepatocyte differentiation, maturation, function and stabilization of phenotype compared to Matrigel cultured cells. Laminin cultured cells arrange themselves in lobule like structures, express MRP1 and MRP2 and are capable of biliary efflux and show significant effect on hepatocyte P450 enzyme metabolic activity. The GMP-ready hepatocyte differentiation protocol is published in Stem Cell Reports.


Dr. Kate Cameron
PostDoc, University of Edinburgh, UK

                                                                                                  2015-04-30

 

 

hESC-derived hepatocyte differentiation on laminin

Neural stem cells in health and disease

Here, Anna presents how studies of neural stem cells and neurons derived from iPS cells of patients show faithful mimicking of known disease phenotypes in our cellular models of disease, like Alzheimer’s disease, autism, and Down syndrome. She has discovered several novel disease specific cellular phenotypes in their in vitro models, many of them involved in accurate performance of the neural progenitors.


Dr. Anna Falk
Ass. Prof., Karolinska Institutet, Sweden                                                                                       

                                                                                                  2015-04-30

 

Neural stem cells on laminin

DNA DAMAGE AND LOW-GRADE MOSAICISM IN HESC CULTURES

Claudia presents data around the frequency and mechanisms behind DNA damage and low-grade mosaicism in hESC cultures. hESC and somatic cell lines frequently acquire segmental copy number variations in the Megabase-scale, forming a non-clonal or low-grade genetic mosaic. The results have been published in Stem Cell Reports showing that hESC grown on laminin-521 show a decreased proneness to acquiring DNA damage during suboptimal culture conditions, such as medium acidification during high culture density.


Dr. Claudia Spits
Ass. Prof., Vrije Universiteit Brussel, Belgium

                                                                                                2015-04-30

 

Genetic stability of human ES cell cultures on laminin

Efficient pancreatic islet maintenance and expansion

Biologically relevant laminins enable mouse pancreatic islets in vitro culture: expansion, phenotype maintenance and glucose-dependent insulin release. When cultured upon selected combination of islet-specific laminins, pancreatic islets exhibited following effects: (1) robust spreading into flattened adherent clusters with heterocellular organization maintaining local cell contacts, (2) islets, cultured upon the selected combination, maintain functional ability for glucose-dependent insulin release, (3) all the endocrine cell types (alpha, beta, delta, PP-cels) maintain specific markers expression (glucagon, insulin/C-peptide/PDX-1, somatostatin, pancreatic polypeptide) and ability to proliferate.

 

Dr. Anna Domogatskaya
Senior Scientist, Karolinska Institute, Sweden

                                                                                                  2014-04-16

 

 

Maintenance and expansion of pancreatic islets on laminin

Clinically compliant hESC-derived RPE cells for advanced macula degeneration

hESC lines derived and cultured on recombinant human laminin 521 under fully chemically-defined and animal substance-free conditions. The differentiated RPE cells exhibit native characteristics including morphology, pigmentation, marker expression, monolayer integrity, polarization and phagocytic activity. The authors are established a large-eyed geographic atrophy model that allowed in vivo imaging of the hESC-RPE and host retina. The protocol for clinical compliant hESC derived RPE cells has been published in Stem Cell Reports.


Dr. Sonya Stenfelt
PostDoc, Karolinska Institute, Sweden

                                                                                                  2014-04-16

 

 

Clinically compliant hESC derived RPE cells

Defined and xeno-free culture of high quality hESC and iPSC

Laminin-521 allow not only self-renewal, but also cloning, derivation, and even clonal derivation of new hES cell lines under defined and xeno-free conditions. Laminin-521 induce fast migration of the cells and laminin isoform specific activation of PI-3K/Akt pathway through interaction with α6β1 integrin. All the hES and human induced pluripotent stem cell lines were confirmed to be genetically stable and pluripotent in in vivo and in vitro assays after extensive culturing on the new substrata. The defined and xeno-free long-term hPSC culture protocol has been published in Nature Communications.


Dr. Sergey Rodin
Senior Researcher, Karolinska Institute, Sweden

                                                                                                  2014-04-16

 

Defined and xeno-free derivation and expansion of hESC and iPSC on laminin-521

Watch as cells grow -
Culture ES and iPS cells on laminin

Stem cell culturing of human ES and iPS cells on recombinant laminin matrices is easy and efficient. Human ES and iPS cells grow as monolayers in a completely defined and xeno-free cell culture environment that enables self-renewal and pluripotency without the use of ROCK inhibitors. Here, we show hES cells plated as single cells on top of recombinant laminin-521, growing from day 0 to day 5 when they are ready to be split again. During the first 1-2 days, a picture was taken every 15 minutes. Days 3-5 a picture was taken every hour.

                                                                                                  2014-04-16

Tissue specific laminins

Laminins are tissue-specific glycoproteins in the basement membrane, important for cell proliferation, migration and differentiation, and tissue development.

Laminin-521 and laminin-511 are expressed by cells in the inner cell mass of the blastocyst and are therefore optimal as substrates for human pluripotent stem cells. Human recombinant laminin isoforms are developed for defined and xeno-free stem cell culturing and primary cell cultures.

                                                                                                  2014-04-16

Stem cell culture on laminin-521

Stem cell culturing is easy on human recombinant Laminin-521 (LN521). It facilitates self-renewal of both pluripotent human embryonic stem cells and induced pluripotent stem cells in a chemically defined, feeder-free and animal-protein free cell culture system.

LN521 allows the survival and expansion of human ES and iPS cells after plating from single cell suspension. LN521 based stem cell cultures grow as monolayers on top of the laminin substrate and remain pluripotent without spontaneous differentiation. To culture embryonic stem cells on LN521 enables easy and efficient expansion of your hESCs and iPSCs and provides a biorelevant in vitro stem cell niche. 

                                                                                                  2014-04-16

 

Biorelevant laminins

Dr. Kristian Tryggvason tells the story about BioLamina and why biologically relevant laminins are the best matrix for cell culturing, including culture of stem cells and primary cells. 

                                                                                                  2014-04-16

Derivation and culture of stem cells and primary cells on specific laminins

Professor Karl Tryggvason discusses the use of biologically relevant human recombinant laminins from BioLamina that successfully recreates specific cell niches in the cell culture dish. This enables robust stem cell culture methods that in practical solves all current technical challenges with culturing human ES and iPS cells. It also describes novel data for culturing primary cells such as cardiomyocytes, pancreatic beta cells and endothelial cells. 

                                                                                                  2014-04-16