BioLamina's posters at ISSCR/StemBANCC/Basel Stem Cell Network 2017 international symposium
Basel, February 27 - March 1:


Abstract ID: P100
Presented on Wednesday, March 1, 2017: 12:30 PM-2:00 PM

CLINICALLY COMPLIANT HUMAN PSC CULTURE CONDITIONS SUPPORT EFFICIENT CLONAL SURVIVAL AND RAPID SCALE-UP

Jesper Ericsson, Yi Sun, Louise Hagbard, Zhijie Xiao and Therese Kallur, BioLamina AB, Sundbyberg, Sweden

 

The lack of defined, xeno-free, easy and robust methods for efficient expansion of human pluripotent stem cells (PSCs) has hindered both the advancement of basic research and human cell therapy, much due to high experimental variation and poor quality cells with phenotypic and genetic changes.

Laminin-521 (LN-521) is a protein naturally expressed by human PSCs and is a critical factor of the pluripotent stem cell niche. Laminins influence adhesion, differentiation, migration, phenotypic stability, anoikis resistance and functionality of all cells associated to it.  

LN-521 is a human and recombinant protein and can easily be used as a cell culture substrate. When used for human PSCs they can be cultured for over 80 single cell passages without any abnormal genetic aberrations and with maintained expression of pluripotency markers. Human PSCs on LN-521 expand twice as fast compared to other matrices and can be split 1:20 or up to 1:30 as single cells without the addition of ROCK inhibitor. Furthermore, LN-521 can be used as microcarrier coating for generating clinically relevant quantities of human PSCs thus offering a scalable and GMP-compatible bioprocessing platform.

Moreover, true clonal culture, important for cell fate tracking, gene function analyses and editing, is possible by using LN-521 in combination with E-cadherin. Human embryonic stem cell (hESC) lines can even be derived from a single blastomere under chemically defined and xeno-free condition on LN-521, thereby circumventing the ethical issues associated with hESCs.

The simplicity and reliability of the culture procedure, the rapid cell amplification and the genetic stability of the cells make LN-521 a suitable as reagent in clinical trials for human PSC-based therapy.

In conclusion, we show that LN-521 is an optimal matrix for human PSC culture due to its biological relevance that allows derivation, clonal cultivation, stable long-term pluripotent cell growth and scalability. The robust method allows minimum culture maintenance and standardized protocols, which can easily be adapted to automation platforms, making LN-521 a suitable reagent choice for human cell therapy trials.

 

Abstract ID: P104
Presented on Wednesday, March 1, 2017: 12:30 PM-2:00 PM

TISSUE SPECIFIC LAMININS GENERATE AUTHENTIC AND CLINICALLY COMPLIANT CELLS

Louise Hagbard, Zhijie Xiao, Therese Kallur, Jesper Ericsson and Yi Sun, BioLamina AB, Sundbyberg, Sweden

 

The lack of defined, xeno-free, robust methods for expansion and specialization of human pluripotent stem cells (hPSC) towards different cell types has hindered both the advancement of basic research and the translation into clinical settings.

The expression and composition of the basement membrane proteins are essential for embryonic morphogenesis and adult tissue functions. Laminins are the only basement membrane proteins that are tissue specific and with the use of the specific combination of xeno-free and defined human recombinant laminins, the natural environment for each specific cell type can be created, generating high quality cell with homogenous phenotypes.

Laminin-521 (LN-521) is the laminin isoform naturally expressed by hPSCs and is also one of the most commonly expressed laminins after birth. LN-521 maintains high degree of hPSC homogeneity, pluripotency and genetic stability when used in vitro.

By using different laminins, differentiation of hPSCs can be made robust, reliable and with increased efficiency. Differentiation of LN-521 cultured hPSCs can be made to:

  • dopaminergic progenitors with LN-111, resulting in an increased yield of >40 times compared to standard EB-based protocols. The GMP-compatible LN-111 based protocol gives a very homogenous population of cells predicted to have a good graft outcome. Due to the high yield the cost of reagent per transplant is very low.
  • RPE cells, exhibiting native characteristics including morphology, pigmentation, marker expression, polarization and phagocytic activity. Transplanted cells exhibit long-term integration and photoreceptor rescue capacity.
  • hepatocytes that in vitro are highly organized, similar to primary tissue, and with significantly increase in metabolic functions.
  • endothelial cells through a defined and xeno-free protocol rendering 95% functional cells facilitating production of stable endothelial cells for vascular disease modeling and treatment.

Laminin coating substrates, compatible with GMP requirements, are being developed which allow the transition of these pre-clinical research protocols into clinical settings.

Other posters to look out for

 

Abstract #: 1102
Time: Wednesday, June 22 at 7:30 - 8:30 pm
Fluid shear stress induces cytochrome activity in 2D- and 3D-cultured human embryonic stem cell-derived hepatocyte-like cells


Abstract #: 2017

Time: Wednesday, June 22 at 6:30 - 7:30 pm
Generation of corneal endothelial cells from human pluripotent stem cells


Abstract #: 2176

Time: Wednesday, June 22 at 7:30 - 8:30 pm
Standardized approaches for evaluation of the definitive endoderm differentiation bias between individual hESC lines


Abstract #: 4020

Time: Wednesday, June 22 at 7:30 - 8:30 pm
Regeneration of myocardial infarcted mouse heart with fully-defined human cardiac progenitor cells


Abstract #: 2015
Time: Thursday, June 23 at 6:00 - 7:00 pm
Tumorigenicity and other safety studies for hESC-derived RPE cells: a step forward in age-related macular degeneration treatment


Abstract #: 3021
 
Time: Thursday, June 23 at 6:00 - 7:00 pm
Recombinant human laminin-521 supports PSC survival in essential 8 media during critical transitions


Abstract #: 3015

Time: Thursday, June 23 at 6:00 - 7:00 pm
GMP compliant stable and efficient expansion of pluripotent stem cells in a closed cultivation system


Abstract #: 3016

Time: Thursday, June 23 at 7:00 - 8:00 pm
Comparison of pluripotent stem cell processes and health when cultured on matrigel, geltrex, and laminin-521 using stemcellQC, a video bioinformatics toolkit

 
Abstract #: 1089
Time: Friday, June 24 at 6:00 - 7:00 pm
Rotator cuff tear state modulates the differentiation and self-renewal of human skeletal muscle progenitor cells


Abstract #: 1097

Time: Friday, June 24 at 6:00 - 7:00 pm
Laminin-based matrices for hepatic-lineage specification of human pluripotent stem cells

Abstract #: 3003
Time: Friday, June 24 at 6:00 - 7:00 pm
Innovative culture of hiPSCs in suspension on alginate-microcarrier


Abstract #: 3022
Time: Friday, June 24 at 7:00 - 8:00 pm
Recombinant laminin-521 precoated cultureware support stem cell culture and differentiation


Abstract #: 4028
Time: Friday, June 24 at 7:00 - 8:00 pm
The effect of surface composition on pluripotency, naive state and efficiency of directed differentiation

BioLamina's posters at ISSCR 2016 Annual Meeting
in San Francisco, June 22 - 25:


Abstract ID: 1981
Presented on Thursday, June 23, 2016: 6:00 PM-7:00 PM

Tissue Specific Laminins Generate Clinically Compliant hPSC Cultures for Clinical Trials 

Louise Hagbard, Zhijie Xiao, Therese Kallur, Jesper Ericsson and Yi Sun, BioLamina AB, Sundbyberg, Sweden


The lack of defined, xeno-free, robust methods for expansion and specialization of human pluripotent stem cells (hPSC) has hindered both the advancement of basic research and the translation into clinical settings. The expression and composition of the basement membrane proteins are essential for embryonic morphogenesis and adult functions. Laminins are the only basement membrane proteins that are tissue specific and with the use of the specific combination of xeno-free and defined human recombinant laminins, the natural environment for each specific cell type can be created, which in turn generate high quality cell cultures of well defined phenotypes. Laminin-521 (LN-521) is the laminin isoform naturally expressed by hPSCs and is also one of the most commonly expressed laminins after birth. LN-521 maintains high degree of hPSC homogeneity, pluripotency and genetic stability when used in vitro. Due to robust support provided by LN-521, human embryonic stem cell (hESC) lines can even be derived from a single blastomere, circumventing the ethical issues associated with hESCs. Here, we show how culture on LN-521, alone or in combination with other laminin isoforms, can been used to generate specialized cells from hPSC. 1) RPE cells can effectively be derived from LN-521 cultured hESC, exhibiting native characteristics including morphology, pigmentation, marker expression, polarization and phagocytic activity. Transplanted cells exhibit long-term integration and photoreceptor rescue capacity. 2) A highly pure population of dopaminergic progenitors can effectively be derived from laminin cultured hESC, with a 30-fold increase in cell yield compared to previous protocols. 3) By using heart specific laminins in the natural combination, high ratio of beating cardiomyocytes with characteristic morphology and marker expression can be generated. 4) Improved hepatocyte specification and maturation of hESC cultured on LN-521 and other liver specific laminin isoforms. The hepatic cells are highly organized, similar to primary tissue, and with significantly increase in metabolic functions. In addition, we also show how a laminin-521 coating substrate, fully compatible with GMP requirements, is being developed which facilitates the transition of these pre-clinical research protocols into clinical settings.

 


Abstract ID:
1965
Presented on Thursday, June 23, 2016: 7:00 PM-8:00 PM

Clinically Compliant Human PSC Culture Conditions Support Efficient Clonal Survival and Rapid Scale-Up 

Jesper Ericsson, Louise Hagbard, Zhijie Xiao, Therese Kallur, and Yi Sun, BioLamina AB, Sundbyberg, Sweden


The lack of defined, xeno-free, easy and robust methods for efficient expansion of human pluripotent stem cells (PSCs) has hindered both the advancement of basic research and human cell therapy, much due to high experimental variation and poor quality cells with phenotypic and genetic changes. Laminin-521 (LN-521) is a protein naturally expressed by human PSCs and is a critical factor of the pluripotent stem cell niche. Laminins influence adhesion, differentiation, migration, phenotypic stability, anoikis resistance and functionality of all cells associated to it. LN-521 is a human and recombinant protein and can easily be used as a cell culture substrate. When used for human PSCs they can be cultured for over 80 single cell passages without any abnormal genetic aberrations and with maintained expression of pluripotency markers. Human PSCs on LN-521 expand twice as fast compared to other matrices and can be split 1:20 or up to 1:30 as single cells without the addition of ROCK inhibitor. Furthermore, LN-521 can be used as microcarrier coating for generating clinically relevant quantities of human PSCs thus offering a scalable and GMP-compatible bioprocessing platform. Moreover, true clonal culture, important for cell fate tracking, gene function analyses and editing, is possible by using LN-521 in combination with E-cadherin. Human embryonic stem cell (hESC) lines can even be derived from a single blastomere under chemically defined and xeno-free condition on LN-521, thereby circumventing the ethical issues associated with hESCs. The simplicity and reliability of the culture procedure, the rapid cell amplification and the genetic stability of the cells make LN-521 a suitable as reagent in clinical trials for human PSC-based therapy. In conclusion, we show that LN-521 is an optimal matrix for human PSC culture due to its biological relevance that allows derivation, clonal cultivation, stable long-term pluripotent cell growth and scalability. The robust method allows minimum culture maintenance and standardized protocols, which can easily be adapted to automation platforms, making LN-521 a suitable reagent choice for human cell therapy trials.

Other posters to look out for

 

Poster # 1135
Time: Wednesday June 24, at 6:30 - 8:30 pm

SELF-ORGANISATION AND DIFFERENTIATION OF HESC DERIVED HEPATOCYTES UNDER DEFINED CONDITIONS

Authors: Cameron, Kate R., Tan, Rosanne, Wang, Yu, Lucendo Villarin, Baltasar, Szkolnicka, Dagmara, Forbes, Stuart J., Hay, David C.
The University of Edinburgh; Edinburgh BioQuarter, Edinburgh, United Kingdom

 

Poster # 1511
Time: Wednesday June 24, at 6:30 - 8:30 pm

OPTIMIZATION OF EMBRYOID BODY FORMATION FROM SINGLE CELL SUSPENSIONS OF HUMAN EMBRYONIC STEM CELLS CULTURED ON LAMININ-521

Authors: Dziedzicka, Dominika, Markouli, Christina, Barbé, Lise, Spits, Claudia, Sermon, Karen, Geens, Mieke, Research Group Reproduction and Genetics, Vrije Universiteit Brussel, Brussels, Belgium

 

Poster # 1521
Time: Wednesday June 24, at 6:30 - :30 pm

STREAMLINING THE PGE IPSC LINE GENERATION - ESTABLISHING AN EFFICIENT AND QUALITATIVE PLATFORM FROM TRANSFECTION TO TARGET CLONE

Authors: Jonebring, Anna Kristina1, Jarl, Lisa1, Boreström, Cecilia1, Engberg, Susanna1, Magnusson, Björn1, Di Castro, Silvio2, Brolén, Gabriella1, 1RAD Discovery Sciences, AstraZeneca, Mölndal, Sweden, 2Screening Sciences - Discovery Sciences, AstraZeneca, Mölndal, Sweden

 

Poster # 1413
Time: Friday June 26, at 6:00 - 8:00 pm

IDENTIFICATION OF CARDIAC PROGENITORS DIFFERENTIATED FROM PLURIPOTENT HUMAN EMBRYONIC STEM CELLS USING MUSCLE-SPECIFIC LAMININ MATRICES

Authors: Yap, Lynn1, Sun, Yi2, Wang, Jiong-Wei3, Ohman, Miina Karelia1, Chai, Xiaoran1, Chong, Liyen1, Cook, Stuart1, Ghosh, Sujoy1, de Kleijn, Dominique P.V.4, Tryggvason, Karl1, 1Duke-NUS Graduate Medical School, Singapore, Singapore, 2BioLamina AB, Stockholm, Sweden, 3National University of Singapore, Singapore, Singapore, 4Utrecht Medical Center, Utrecht, Netherlands

 

Poster # 1521
Time: Friday June 26, at 6:00 - 8:00 pm

HUMAN PLURIPOTENT STEM CELL GROWTH ON LAMININ 521 COATED, MICROCARRIER CULTURES

Authors: Oh, Steve, Chen, Allen, Lam, Alan, Reuveny, Shaul Bioprocessing Technology Inst, Singapore, Singapore

Poster #: 1500
When: Wednesday June 24, at 6:30 - 8:30 pm

HEART, BRAIN, EYE AND PANCREAS CELLS THRIVE ON BIOLOGICALLY RELEVANT

DEFINED AND XENO-FREE LAMININS 

Authors: Kallur, Therese, Ericsson, Jesper, BioLamina AB, Stockholm, Sweden


Abstract

Laminins are a group of 16 heterotrimeric glycoprotein isoforms found in the basement membrane in the extracellular matrix and are composed of α, β and γ chains, and are the only tissue-specific proteins in the basement membrane. They are thus critical factors differentiating one cell niche from another and influence the behavior of associated cells, such as adhesion, differentiation, migration, phenotype stability, and resistance to anoikis.

The use of specific laminins for tissue culture and cell therapy applications have been hampered by lack of access to most laminin isoforms. Tissue-purified laminins have been available for years but often result in poor quality due to protein degradation, lot-to-lot variation and impurities, resulting in variable and unreliable research results. We have now solved these problems by successful production of human recombinant laminins and have shown that individual laminin isoforms drastically improve the functional properties of different cells.

Pluripotent stem cells: By using LN-521, which is naturally expressed by human PSCs, we can culture stem cells for over 130 single cell passages at split ratios of 1:10-1:30, without any abnormal genetic aberrations and with maintained expression of pluripotency markers.
Cardiomyocytes: By using heart specific laminins, LN-211, LN-221 and LN-521 in the natural combination of adult heart cell expression, the differentiation can now be fully controlled in a defined and xeno-free context and the number of hPSC-derived beating cardiomyocytes is significantly increased.
Neurons and glia: Neural stem cells prefer LN-521, different neuronal subtypes require primarily LN-511/521 and LN-111 for full maturation, and glia cells express LN-111, LN-211 and LN521.
RPE cells: Robust retinal pigmented epithelium cell culture has been reported on LN-521/511, LN-111 and LN-332.

In conclusion, cell culture of primary cells and stem cells is reliable and robust when growing cells on the natural human recombinant laminin that match the in vivo niche. Almost all cells grow on specific laminins in the human body and as they now are available as recombinant laminins it makes cell culture in a physiologically relevant environment possible, making production of clinically relevant cells possible

 

 

Poster #: 1531
When: Friday June 26, at 6:00 - 8:00 pm

SCALABLE MONOLAYER PSC CULTURE ON LN521 WITH ROBUST SINGLE CELL

PASSAGE AND FREE WEEKENDS UNDER XENO-FREE AND DEFINED CONDITIONS

Authors: Xiao, Zhijie, Ericsson, Jesper, Hagbard, Louise, Sun, Yi, Kallur, Therese 


Abstract

he lack of defined, xeno-free, easy and robust methods for efficient expansion of human pluripotent stem cells (hPSC) has hindered both the advancement of basic research, due to high experimental variation and poor quality cells with phenotypic and genetic changes, and human cell therapy requiring absolute safe methods and large numbers of low passage cells.

By using a human recombinant protein naturally expressed by hPSCs, LN-521, we can culture hPSCs for over 80 single cell passages without any abnormal genetic aberrations and with maintained expression of pluripotency markers. Cells cultured on LN-521 grow twice as fast compared to other matrices and can be split 1:20 or up to 1:30 as single cells without the addition of artificial ROCK inhibitor (Rodin, Nat Comm 2014). The simplicity and reliability of the procedure, speed of cell amplification and the genetic stability of the cells make LN-521 suitable as reagent in clinical trials for PSC-based therapy.

Furthermore, true clonal growth, important for cell fate tracking, gene function analyses and editing, without inhibitors of anoikis, is possible by using LN-521 and E-cadherin (Rodin, Nat Prot 2014). The same authors also demonstrated chemically defined and xeno-free derivation of new clinical hESC lines from single blastomers. In essence, circumventing the ethical dilemma of destroying the surplus embryos donated by couples going through fertility treatments.

LN-521 provides a biorelevant niche for hPSCs and we are now able to show that hPSCs can be cultured without the need of daily feeding. Data from 10 consecutive passages on three different lines reveal no significant differences in cell morphology, proliferation or the expression of pluripotency markers such as Oct4 and Nanog. The results show that the expression levels are similar to both the daily fed group and starting samples. Chromosome analysis after 10 passages indicated normal karyotype for all groups.

In conclusion, we show that LN-521 is an optimal matrix for hPSC culture due to its biological relevance allowing derivation, clonal cultivation and robust long-term pluripotent cell growth. The robust method allows minimum culture maintenance and standardized protocols, which can easily be adapted to automation platforms, making LN-521 a suitable reagent choice for human cell therapy trials.

EFFICIENT CRYOPRESERVATION

Efficient cryopreservation of human PSCs in a chemically defined cGMP produced, serum-, xeno- and DMSO-free freezing medium 

PRINT POSTER  

 

WEEKEND-FREE CULTURE

Reduced or no feeding procedure for human PSCs on LN-521 stem cell matrix

PRINT POSTER

 

STEM CELL CULTURE MADE EASY

LN-521TM enables derivation, clonal culture and easy single- cell passage of human pluripotent stem cells without ROCKi

PRINT POSTER