Development and application of ec23®

Development and application of the

Synthetic Retinoid ec23®

and

Cell Differentiation

AMSBIO is the global source for ec23®.

ec23® is a registered trade mark of and manufactured by Reinnervate.

Stem Cells and Cell Differentiation

Differentiated tissues

Stem cells

Cell division

Ste

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ell

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ec23®

Highly controlled process

Regulated by specific molecular pathways

Differentiation

Neurogenesis in the Embryo

Differentiated

Neural cell

Postmitotic

Neuroblast

Ventricular

Highly regulated process involving several well characterised

signalling pathways including : Wnt; SHH; FGF; RA; BMP; etc.

Proliferating

Neuroprogenitors

Ventricular

Zone

Neural stem cell

ec23®

Neurogenesis by Cultured Human Stem Cells

Stem cells

Example applications:

� Basic research

� Development of pharmaceuticals

� Disease modelling

Neural Tissues

�

� Toxicological testing

ec23®

Development of Synthetic Retinoids That Enhance Cell Differentiation

OH

O

all-transretinoic acid

(ATRA)

• ATRA is well known for its ability to modulate cell function and neural development

• However, naturally occurring retinoids such as ATRA are unstable and degrade readily during use in the cell culture laboratory

Molecular modelling

Characterisation& purity testing

Chemicalsynthesis

Biologicalscreening

• We have developed a series of synthetic retinoids that maintain chemical stability and possess biological activity equivalent to ATRA

• Such degradation has been shown to result in variable induction of cell differentiation

ec23®

Molecular Modeling of Key Compounds

CO2H

ATRA

CO2Hec23®

ec23®

Stability of ATRA and Synthetic Retinoid ec23®

Compound analysis by NMR

ec23®ATRA

• ATRA is light and temperature sensitive and readily isomerises

• ec23® remains stable over prolonged periods of intense li ght exposure

¹H NMR (500 MHz) spectrum (δ 5.40-7.80) of ATRA in D16-DMSO in a glass NMR tube after 3 days in the dark under air (top), versus 3 days exposure to fluorescent light (bottom ).

¹H NMR (500 MHz) spectrum (δ 7.20-8.10) of EC23® in D16-DMSO in a glass NMR tube after 3 days in the dark under air (top), versus 3 days exposure to fluorescent light (bottom).

ec23®

Robust Induction of Neurogenesis

1 µM ATRA

� Induction of alternative lineages

20

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ns

1 µM ec23®

� Enhanced neurogenesis

� Decreased heterogeneity

ec23®

ec23®ATRA

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Stem Cell Derived Neurons Show Enhanced Neuritogenesis

Example shows significant increase in neurite outgrowth when treated with synthetic retinoid ec23®

1µM ATRA 1µM ec23®

ec23®

Optimizing Protocols to Deliver Enhanced Neurogenesis

• Induction of neurogenesis and

formation of neurites by

synthetic retinoid ec23®

• Optimised protocols enable

enhanced neurogenesis by

human stem cellshuman stem cells

• Applications for basic research,

screening, toxicological testing

and disease modelling

Human stem cell derived neurons induced by synthetic retinoid ec23®

ec23®

Enhancing the Epidermal Barrier by Retinoid Treatment

• Retinoid treatment inhibits differentiation and promotes proliferation of epidermal cells(Fuchs and Green, 1981)

• ec23® is a stable synthetic retinoid designed to mimic biological activity of all-trans-retinoic acid

• Initial exposure of keratinocytes to ec23® is predicted to thicken the epidermal barrier prior to differentiation at the air/liquid interface

brightfield

involucrinstaining

control 1µM ec23®

ec23®

Epidermal Thickening In Vivo

A

epi

der

hyp

B C

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Topical treatment of skin with ec23® results in a ~3-fold thickening of the adult mouse epidermis

Applications in basic research, disease modelling and potential therapeutic / cosmetic use

Control ATRA 50µM ec23® 50µM

ec23®

Summary

• ec23® is a stable derivative of ATRA and mimics its biological activity

• ec23® is 1-2 orders of magnitude more potent that ATRA

• ec23® can be used either directly or as a supplement to existing reagents• ec23® can be used either directly or as a supplement to existing reagents

• ec23® is versatile and has multiple applications in modulating cell growth

ec23®

Main publications

� Retinoid supplementation of differentiating human neural progenitors and embryonic stem cells leads to enhanced neurogenesis in vitro.

Christie VB, Maltman DJ, Henderson AP, Whiting A, Marder TB, Lako M, Przyborski SA.

J Neurosci Methods. 2010 Nov 30;193(2):239-45.

� Synthetic retinoids: structure-activity relationships.

Barnard JH, Collings JC, Whiting A, Przyborski SA, Marder TB.Barnard JH, Collings JC, Whiting A, Przyborski SA, Marder TB.

Chemistry. 2009 Nov 2;15(43):11430-42.

� Proteomic profiling of the stem cell response to retinoic acid and synthetic retinoid analogues: identification of major retinoid-inducible proteins.

Maltman DJ, Christie VB, Collings JC, Barnard JH, Fenyk S, Marder TB, Whiting A, Przyborski SA.

Mol Biosyst. 2009 May;5(5):458-71.

ec23®

Further information

Contact Information

AMS Biotechnology

184 Milton Park, Abingdon, Oxon, OX14 4SEinfo@amsbio.com

+49 (0) 69 77 9099+44 (0) 1235 828200

info@amsbio.com

www.amsbio.com

+41 (0) 91 604 5522+1 949 265 7717

Your source for life science products & services

AMSBIO is the global source for ec23®.

ec23® is a registered trade mark of and manufactured by Reinnervate.

+1 800 987 0985 (Toll Free)