Técnicas en biología molecular y celular. In: Iwasa J, Marshall W. eds. Biología Celular y Molecular. Conceptos y experimentos, 8e New York, NY: McGraw-Hill; . http://accessmedicina.mhmedical.com.consultaremota.upb.edu.co/content.aspx?bookid=2817&sectionid=239341463. Accessed marzo 04, 2020.

1. This is your
presentation title
ANA MARÍA AGUDELO HERRERA
THIRD SEMESTER- MEDICINE. 2020-1

2. INTRODUCTION
TRANSFECTION
Deliver nucleotic
acid into an
eykariotic cell
Electroporation
Lipofection
Calcium phosfate or
DEAE-dextrano

3. INTRODUCTION
PROTEIN
CORONA
Interaction between the
GOCL/DNA
(grapholipoplexes) and
biological fluid such as
human plasma
Complex exposed to
proteins get
envolved by a layer
called PROTEIN
CORONA.
The layer evolves with
protein concentration,
leading to biocoronated
with modified
physicochemical properties

4. OBJECTIVE
‐ Explore the effect of the protein
corona on the transfection
efficiency of lipid-coated Graphene
cell transfection reagents.

5. METHODS
PREPARATION OF GOCL NANOPARTICLES
FUNDAMENTS FOR WHAT PURPOSE
To obtain an homogenous
solution of graphene oxide
(GO)- cationic lipid (GOCL
nanoparticles.
The NanoAssemblrTM Benchtop employs a
microfluidic cartridge and pumps two
different solutions into two microfluidic
channels that converge in a terminal
channel by fine‐tuning mixing parameters
as, Total Flow Rate (TFR) and Flow Rate
Ratio (FRR).

6. METHODS
CELL CULTURE
FUNDAMENTS FOR WHAT PURPOSE
Breast and preserve
cancer cell lines,
glioblastoma cell line
and colorectal
adenocarcinoma cell
line.
MDA‐MB, MCF‐7 and U‐87 cells
were preserved in DMEM
supplemented with 10% fetal
bovine serum (FBS) and CACO‐2
cells were preserved in RPMI
medium supplemented with 20% FBS.

7. METHODS
TRANSFECTION EFFICIENCY EXPERIMENTS
FUNDAMENTS FOR WHAT PURPOSE
To carry out the
transfection, analyse the
luminerase expression and
determine the transfection
efficency.
Treat the cell lines MDA‐MB,
MCF‐7, U‐87 MG and CACO‐2
with the GOCL/DNA complex.

8. METHODS
FUNDAMENTS FOR WHAT PURPOSE
1D SDS PAGE GEL ELECTROPHORESIS
For the protein corona
characterisation.
Is a method that separates protein by
molecular weight. In this method,
samples are weighed and dissolved in
sodium dodecyl sulfate (SDS). SDS is
a negatively charged detergent that
has both hydrophilic and hydrophobic
regions.

9. RESULTS
‐ Protein Corona
composition of both
systems was
influenced by
protein
concentration

10. RESULTS
‐ At low protein concentration
(HP < 5%), protein patterns of
GOCL NPs and grapholipoplexes
were almost superimposable.
‐ At high protein concentration
(HP < 20%), marked differences
in the protein profiles of GOCL
NPs and grapholipoplexes were
found.
- Low protein
concentration, they
preferentially bind
to the lipid surface
of grapholipoplexes.

11. RESULTS
MDA‐MB cells treated
with fluorescently
labelled (green)
pristine and
biocoronated
grapholipoplexes (HP =
20%)
Cells were stained with
LysoTracker Deep Red
(red), a lysosomal
marker, and Dapi to
stain the nuclei.

12. RESULTS
‐ Grapholipoplexes
colocalising with
lysosomes gave rise to
yellow clusters
Pristin grapholipoplexes
Bicoronated counterpart
VS
“It has been demonstrated the
formation of PC provides a
capacity to alter both cellular
and intracellular location of
nanoparticles”

13. Disscussion
Vestibulum congueVestibulum congueVestibulum congueVestibulum congue
AUTHOR HYPOTHESIS YES/NO
Giulimondi,
F.; et al.
“Evolution profiles of size and zeta‐potential of
biocoronated grapholipoplexes reported in Figure 2 closely
resemble those previously reported for cationic
liposome‐protein complexes”
Bertoli, F.;
et al
“In this view, PC formed in human plasma (e.g., upon
systemic administration) may compromise endosomal escape of
biocoronated grapholipoplexes, shuttling them to lysosomal
compartments”
Bigdeli, A.;
et al
“This inversion in surface charge is likely to be
responsible for reduced electrostatic interaction with cancer
cells that, in turn, may lead to lower cellular uptake.”

14. “CONCLUSIONS
To conclude, the layer of protein corona formed by a high
concentration of proteins can reduce the transfection efficiency in
the cancer line cells used in the experiment and also can increase the
probability to digest of the DNA useful load at the intracellular
level.
In conlcusion if we expose grapholipoplexes complex to an artificial
coronad formed by a low concentration of proteins, can increase the
transfection efficiency and low citotoxicity.