The development of in vitro models of BBB has
yielded important insights into our understanding
of the pathophysiology of the BBB. These
models have progressed from simple monocultures
of endothelial cells to more complex
coculture systems in which endothelial cells are
grown on porous cell culture membranes and
cocultured with astrocytes .
Human endothelial cells (HECs) from the
umbilical vein (American Type Culture Collection)
were cultured in M199 containing 10%
dyalised fetal bovine serum (FBS), 1 mmol/L
glutamine, Endothelial Cell Growth Supplement
(ECGS, 150 mg/mL), 1 mmol/L sodium pyruvate,
and heparin (5 units/mL) . Rat Brain
Microvascular Endothelial Cells (RBMEC)
(Innoprot, Derio, Bizkaia, Spain) were cultured
in Endothelial Cell medium (ECM) supplemented
with 5% dyalised FBS and ECGS. Rat
cortical Astrocytes (RCA) (Innoprot, Derio,
Bizkaia, Spain) and Human Astrocytes (HA)
(ScienCell Carlsbad, CA, USA) were cultured in
Astrocyte medium (AM) containing 5% dyalised
FBS and 1% of Astrocyte Growth Supplement.
Different cocultures were developed i) RBMEC/
RCA and ii) HECs/HA  and defined rat BBB
and human BBB, respectively. The BBB in vitro
models were established using the Transwell
system (Corning, Corning NY, USA) with a
0.4 mm pore size. RCAs or HA (35,000/cm2) were
seeded on the underside of the insert precoated
with Poly-L-lysine (100 mg/mL). Once they
reached confluence, HECs or RBMECs were
seeded on the upper side of the membranes
(60,000/cm2) precoated with fibronectin (50 mg/
mL). BBB was validated by measuring the
transmonolayer electrical resistance (TEER) of
the endothelial monolayer at various time points
using an EndOhm (World Precision Instruments,
Friedberg, Germany). The experiments
were repeated 3 times, each sample being
replicated 3 times. Results were expressed in
Wxcm2  and are presented as the mean S.
Yes, I get that too, but I put it on my forearms. Shortly after I can taste it in my mouth slightly too.
I've been taking Magnoil for a little over a week now; a full dose divided between early a.m. and early evening and have experienced the following effects:
- improved glucose control. It has eliminated the reactive hypoglycemia I almost always experience after lunch and dinner.
- no more stiff legs. Over the past 10 months or so I've had constant really stiff legs, mostly around the hamstring area and sometimes my thighs too. I've tried Mag chloride spray, which helped some near the surface but not deep into the legs, and Magnoil has completely eliminated all stiffness. I can do leg stretches much more easily now.
- less thirst. Sometimes my thirst is insatiable, but since starting Magnoil I don't feel the need to drink nearly as much water.