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PRODUCT RECOMMENDATION

LAMININ-111 and LAMININ-521

 
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PROTOCOL

Generation of high-purity human ventral midbrain dopaminergic progenitors for in vitro maturation and intracerebral transplantation

Highly pure midbrain dopamine cultures under defined and xeno-free conditions 

 


VIEW ON DEMAND

Effective GMP-compliant production of hESC-derived DA neurons on laminin 

Watch webinar on generation of clinically compliant stem cell derived dopaminergic neurons on laminin-111

 


APPLICATION NOTE

Get the dopaminergic neuron application note here
  


CUSTOMER TESTIMONIALS

"We have struggled for a number of years to establish a GMP manufacturing process for generating DA neurons from human embryonic stem cells in order to develop a stem cell based therapy for Parkinson’s Disease. The recombinant laminins from Biolamina really made a world of difference! We now routinely use LN521 and EDTA for passaging the pluripotent stem cells and we have a greater efficiency than ever before. It also turns out that the DA progenitor cells love to grow on LN111 and after introducing this into our protocol we now have a high yield, high quality GMP differentiation ready to use”

                                               Prof. Malin Parmar, Lund University, Sweden

         
More laminin product testimonials here
 


REFERENCES

HOW TO CULTURE DOPAMINERGIC NEURONS ON LAMININ SUBSTRATES

hPSC-derived clinically compliant dopaminergic neuron differentiation and culture on defined human recombinant laminin-521 and laminin-111 cell culture reagentsLaminin culture supports GMP-compatible differentiation of dopaminergic cells

Parmar and colleagues show in Cell Stem Cell (2016) that laminin-111 can be used for the GMP-compatible differentiation of hESCs to dopaminergic cells (Kirkeby, 2016). Creating not only a defined, xeno-free and clinically compliant protocol but also generates a homogenous progenitor population composed of cells expressing the markers identifying right type of mesDA cells. Importantly, the yield of these correctly specified progenitors increases 43-fold compared to other EB-based protocols. The protocol has been validated in several independent labs and with many different hPSC lines both, hESC and hiPSCs. In all cases the hPSCs were expanded using laminin-521 (LN-521), making the starting hPSC population homogenous and stable befor initiation of DA differentiation experiments.  

The authors show how to precisely set the balance of patterning factors to obtain specifically the caudal VM progenitors that give rise to DA-rich grafts. The timely addition of carefully titrated FGF8 to the adherent differentiation protocol, ensures that progenitors of caudal mesencephalic fate is enriched and contaminating progenitors destined for other fates such as neurons in the STN are reduced in numbers. Furthermore, using a panel of PCR markers, developed by using data from this article, batches of progenitors for transplantation can be predictive as giving high numbers of DA neurons after grafting and thus giving better improvement of PD related deficits (Kirkeby, 2016). 

A detailed protocol for better purity and high yield of dopaminergic progenitors for transplantation

The same group recently published a detailed 16-d protocol (Nature Protocols) for obtaining high-purity ventral midbrain (VM) dopamine (DA) progenitors for intracerebral transplantation into animal models and for in vitro maturation into neurons (Nolbrant, 2017). The protocol is free of xeno-derived products and can be performed under good manufacturing practice (GMP) conditions. The authors have successfully transplanted such cells into the rat and the protocol is designed to also be compatible with clinical transplantation into humans. By specifying how to perform quality control (QC), troubleshooting and adaptation of the procedure, this protocol will facilitate implementation in different laboratories and with a variety of hPSC lines. To facilitate reproducibility of experiments and enable shipping of cells between centers, the authors present a method for cryopreservation of the progenitors for subsequent direct transplantation or terminal differentiation into DA neurons (Nolbrant, 2017). 
 

Efficient midbrain dopamine differentiation and improved grafting outcomes in Parkinsonian rodents

In a publication by Niclis et al., the authors report the generation of highly pure midbrain dopamine cultures under feeder-free, fully defined, and xeno-free conditions from hPSCs on laminin-521. The cells display gene, protein, and electrophysiological properties of midbrain neurons, as well as improved grafting outcomes in Parkinsonian rodents. Furthermore, ventral midbrain dopamine neurons were amenable to scalability and cryopreservation, crucial steps for the advancement of cell replacement therapy in Parkinson's disease (Niclis, 2016).

Lu and colleagues show that direct differentiation of dopaminergic neurons also can be performed on laminin-521. The authors reprogrammed iPS cells directly on laminin-521 and then differentiated the cells through an adherent 2D differentiation protocol to DA neurons on laminin-521 (Lu, 2014).

LAMININ KEY ADVANTAGES

  • Clinically compliant differentiation protocol based on the use of laminin-111

  • Using laminin reduces experimental variation

  • >40-fold DA progenitor yield increase on laminin-111

  • Laminin-111 differentiated progenitors have a homogneous expression profile

  • Contaminating PSCs are effectively removed using laminin-111 for 16 days of differentiation

  • Robust protocol replicated with >10 human PSC lines in diff erent laboratories world wide

  • Highly pure midbrain dopamine cultures on laminin-521

  • Differentiation of dopaminergic neurons can be performed on laminin-521

  • Defined and xeno-free culture method

  • Significant reduction of costs due to differentiation efficiency and phenotype authenticity

  • x
  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Neural cells

  • Skeletal muscle cells

  • Kidney cells

  • Hepatic cells

  • Cancer cells

  • Lung cells

  • Intestinal cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Normal and cancerous mammary cells

  • Epithelial cells

Laminin-521

Laminin-521 human stem cell matrix makes pluripotent stem cell culture easy in a defined, xeno-free and biologically relevant cell culture system.

  • x
  • Neural cells

  • Hepatic cells

  • Lung cells

  • Intestinal cells

  • Normal and cancerous mammary cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Skeletal muscle cells

  • Kidney cells

  • Cancer cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Epithelial cells

Laminin-111

Laminin-111 is important during early epithelial development and differentiation of the epiblast, and is commonly used as a general attachment protein in cell culture.

  • x
  • Neural cells

  • Eye cells

  • Endothelial cells

  • Animal stem cells

  • Pancreatic cells

  • Epithelial cells

  • Cardiac cells

  • Cancer cells

  • Hepatic cells

  • Intestinal cells

  • Kidney cells

  • Lung cells

  • Skeletal muscle cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Normal and cancerous mammary cells

LAMscreen

LAMscreen is a kit with all our currently available laminin isoforms so that you can easily screen the best laminins for different cell types.