macroalgae cell culture: an innovative approach to...

Macroalgae cell culture: an

innovative approach to develop

sustainable marine active ingredients

13/12/2016

- Erwan Le Gélébart -

7815 GB 01 November 2016 / An innovative approach to develop sustainable

marine active ingredients from macroalgae



SEPPICIngredients that inspire

7815 GB 01 November 2016



PROFILE

Air Liquide

World leader in gases, technologies and

services for Industry & Health

SEPPIC

is a designer of specialty ingredients

for health & beauty care

BiotechMarine

is one of the world leading specialist

in high-tech bioactive natural substances

extracted from macroalgae & halophyte plants



BiotechMarineSustainable ingredients from the ocean

Bréhat

Archipelago

Pioneer in marine cosmetics in macroalgae &

Seashore plants

extraction

28yearsexpertise

in plant cell culture

with CELTOSOMETM

10yearsexperience

More than

Launch of a new generation of Active Ingredients

From macroalgae cell culture:In

2015CELEBRITYTM

Keypoints:

CELEBRITYTM

An innovative approach

to develop sustainable marine

active ingredients





WORLD’S OCEANS BIODIVERSITY:

a significant source of new active ingredients

Marine flora of the north coast of Brittany (France)

is particularly abundant & diverse, due to favorable

environmental & geographical conditions.

In the last decades, microalgae have been of

growing interest whereas macroalgae stayed

relatively unexploited regarding the large

number of species available.



WHY VALORISING THESE MACROALGAE?

Macroalgae little known:• No traditional use

• No predictable biological activity

• Phytochemistry not described

• No cultivation technique

Some of these algae are not identified

We have seen opportunities

in these difficulties…

Major innovation potential!



• develop a general technique to cultivatethese macroalgae or their cells,

• produce large amount of biomass,

• develop extraction methods,in order to maximise valorisation,

• discover biological activities in biomass extracts.

As it is not possible to harvest these species, we needed to:

VALORISING THESE MACROALGAE:which challenges?

Most of these algae are not available in large quantities & hardly harvestable:

species poorly known rare (biomass) small size hardly identifiable



CELEBRITYTM / 2016 Launch: CONTACTICEL™Urban-life dermo-purifier

CONTACTICEL™, extracted form Acrochaetium

moniliforme an epiphytic macroalgae

CONTACTICEL™ for regulation of oily skin

exposed to pollution

In vitro: • Regulation of the effects of urban dusts on sebocytes

• Reduction of the production of lipids: -17%

Clinical results in a polluted atmosphere: • Visible sebum regulation

• Reduction of anti-pollution marker:

- 61% ozonolyzed squalene



SOURCING EPIPHYTES:a niche of diversity

Same kind of structure observed underwater,

niches that are not explored, containing

small & rare species!

• Plants living at the surface of other plants

• Commonly observed in rainforests

• Contibute to the complexity, structure

& function of the canopy

• Create niches of biodiversity

Epiphytism:

… where 70% of orchids live

as epiphytes in tree canopies

Example of

Amazonian

forest…

Source/©: www.brettcolephotography.com/?search=epiphytic

Gravendeel et al. 2004

http://www.brettcolephotography.com/?search=epiphytic



EPIPHYTES:Expectation of original metabolism

Host (algae)

Biofilm

Epiphytes

Decreasing

light / gaz / nutrient

Excretion of wastes

& secondary Metabolites

Selection of an epiphyte

Close relation between

the host & the epiphyte

Exposed to wastes

& secondary metabolites

High potential of

compound discovery



Acrochaetium moniliforme:an underwater epiphyte/epizoic algae

Source: MARINE

MAMMAL SCIENCE,

32(2): 753–764

(April 2016)

Acrochaetium species: Known to be an epiphyte algaeClose relation between the host & the epiphyte

has it has been described between

Gracilaria chilensis & Acrochaetium sp.

Weinberger et al., 2007

Known to be an epizoic algaeSurprisingly capable to attach & grow

on an highly active Sea Otters

& unexpectedly doesn’t affect the thermal insulation

of the fur which plays a critical role in otter

thermoregulation.

Bentall et al., 2016

Red algae Florideophyceae

30µm < Size of specimen < Few cm

Settles as secondary occupant

of biofilm formed by bacteria

at the surface of hosts



FROM ISOLATION

TO INDUSTRIAL BIOMASS PRODUCTION



FROM BIOMASS TO A PROMISING EXTRACT



Study

synopsis

Standardised

urban dust(SRM1649b - National Institute

of Standards &

Technology/NIST- USA)

tipify atmospheric particulate

matter obtained from an

urban area

Particle size:

● 10%: < 1.5 µm, 90%: < 43 µm

Composition of the urban dust

● Polycyclic aromatic hydrocarbons (PAHs)

● Nitro-substituted PAHs (nitro-PAHs)

● Polychlorinated biphenyl (PCB) congeners

● Chlorinated pesticides

● Inorganic constituents in atmospheric

particulate material (including cadmium)

● other

Evaluation of biological parameters:

•Viability: DNA labelling (Hoechst method)

•Lipids synthesis: neutral lipids production (Bodipy® fluorescent probe labelling)

► Microscope acquisition & image analysis (number of cells & Bodipy® fluorescence

intensity → normalization of the fluorescence / cell).

ANTI-POLLUTIONIN VITRO: Regulation of the effects of urban dust on sebocytes(Innovative in vitro model)



ANTI-POLLUTIONIN VITRO: Regulation of the effects of urban dust on sebocytes(Innovative in vitro model)

Study

results

Urban dust

increase in lipids

production (+40%)

CONTACTICEL™

60% reduction of this

overproduction

Effect not observed

without pollution stress

(data not shown)

suggesting a

mecanism of action

specific to pollution

context.



Study

synopsis

Squalene Ozonolysed squalene

From Synelvia

• Study in Shanghai, China

• 2 groups of 20 women, from 25 to 60-years old, presenting an oily skin

• 1% CONTACTICEL™ formulation vs placebo, twice daily application

• D0 & D56 → sebum collection (forehead) on a specific inert matrix and GC-

MS analysis & quantification:

o Total sebum (neutral lipids)

o Ozonolysed squalene : C27-Pentaenal, C22-Tetraenal and C17-Trienal

(indicators of pollution impact) – only for CONTACTICEL™.

Moderate to Unhealthy

pollution level

(Air Quality Index)PM2.5 & ozone

monitoring

VISIBLE BENEFITS FOR THE SKININ VIVO: Regulation of ozonolysed squalene & sebum excretion



Study

results

• Decrease in sebum (neutral lipids)

• Reduction of pollution impact (ozonolysed squalene)

Improvement of oily skin condition.

D0

D56

VISIBLE BENEFITS FOR THE SKININ VIVO: Regulation of ozonolysed squalene & sebum excretion

D56



CELEBRITYTM

An innovative approach to develop sustainable


Benefits of this approach:

Accessing an entire segment of

biodiversity which remains unexplored

Sustainability: only 1 specimen from

nature allows us to produce biomass

Producing large amount of biomass

from algae not harvestable in nature

Obtaining unialgal biomass

Producing very innovative active

ingredients

Thank you

for your attention!

Contacts

Nota

The analytical specifications warranted are only those mentioned on the certificate of analysis supplied with each delivery of the product.

Except as set forth above, SEPPIC* makes no warranties, whether express, implied or statutory, as to the product which is the subject of this document. Without limiting the generality

of the foregoing, SEPPIC* makes no warranty of merchantability of the product or of the fitness of the product for any particular purpose. Buyer assumes all risk and liability resulting

from the use or sale of the product, whether singly or in combination with other goods. The information set forth herein is furnished free of charge and is based on technical data that

SEPPIC* believes to be reliable. It is intended for use by persons having technical skill and at their own discretion and risk. Since conditions of use are outside SEPPIC*'s control,

SEPPIC* makes no warranties, express or implied, and assumes no liability in connection with any use of this information. Nothing herein is to be taken as a license to operate under

or a recommendation to infringe any patents.

* SEPPIC being:

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ANNEXE

Bibliographic References

Bentall G B, Rosen B H, Kunz J M, Miller M A, Saunders G W and LaRoche N L,

Characterization of the putatively introduced red alga Acrochaetium secundatum

(Acrochaetiales, Rhodophyta) growing epizoically on the pelage of southern sea

otters (Enhydra lutris nereis), Marine Mammal Science, 2016, 32(2):753–764

Gravendeel B, Smithson A, Slik FJW and Schuiteman A, Epiphytism and pollinator

specialization: drivers for orchid diversity? Philosophical Transactions of the Royal

Society B: Biological Sciences, 2004, 359:1523-1535.

Weinberger F, Beltran J, Correa J A., Lion U, Pohnert G and Kumar N, Spore

release in Acrochaetium sp (Rhodophyta) is bacterially controlled, Journal of

Phycology, 2007, 43:235–241



ANNEXE

relative to slide n°11

Characterization of the putatively

introduced red alga Acrochaetium

secundatum (Acrochaetiales,

Rhodophyta) growing epizoically on

the pelage of southern sea otters

(Enhydra lutris nereis)

MARINE MAMMAL SCIENCE,

32(2): 753–764

(April 2016)

GENA B. BENTALL,1,2 U.S. Geological Survey, Western

Ecological Research Center, Santa

Cruz Field Station, 100 Shaffer Road, Santa Cruz,

California 95060, U.S.A.; BARRY H.

ROSEN, U.S. Geological Survey, 12703 Research

Parkway, Orlando, Florida 32826, U.S.A.;

JESSICA M. KUNZ and MELISSA A. MILLER, California

Department of Fish and Wildlife,

Marine Wildlife Veterinary Care and Research Center, 1451

Shaffer Road, Santa Cruz, California

95060, U.S.A.; GARY W. SAUNDERS, University of New

Brunswick PO Box 4400, Fredericton,

New Brunswick E3B 5A3, Canada; NICOLE L.

LAROCHE,3 U.S. Geological Survey,

Western Ecological Research Center, Santa Cruz Field

Station, 100 Shaffer Road, Santa Cruz,

California 95060, U.S.A.

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