Granted Patent B2

US 12,195,755 · App. 16/822,994 · Granted Jan 14, 2025

Inventors: Anita Shukla (Providence, RI); Christina Bailey-Hytholt (Providence, RI); Anubhav Tripathi (Northborough, MA)

Assignee: BROWN UNIVERSITY

C12N5/0605A61K9/1617A61K35/50A61K49/1839C07K14/4715G01N33/5014C12N2506/025G01N2405/04

Abstract

Claims (17)

1. An in vitro, cell-free, human placenta model comprising:

1) a membrane supporting a lipid bilayer, the bilayer including a composition of synthetic lipids and/or natural lipids representing a ratio of lipids measured from a lipid extraction from a human placental cell membrane at a first, second, third trimester, or a term pregnancy;

2) a donor compartment disposed on one side of the bilayer; said compartment holding a test compound in contact with the one side of the bilayer; and

3) a detector for detecting at least one of the test compound's permeation through the bilayer, the test compound's adsorption on the bilayer, or a conductivity effect on the bilayer due to the contact with the test compound; said detecting during and/or after a time that the test compound is in contact with the bilayer;

wherein the in vitro placenta model is capable of simulating the test compound's permeation through, the test compound's adsorption on, or the test compound's conductivity effects on an in vivo human placenta because the ratio of lipids of the in vitro bilayer is measured from the lipid extraction from the human placental cell membrane.

2. The in vitro, cell-free placenta model of claim 1 ,

wherein the lipid bilayer is further comprising:

synthetic lipid vesicles or both synthetic lipid vesicles and synthetic lipid bilayer; and

is further comprising lipids that are the most abundant measured from the human placenta including phosphatidylcholine (PC), phosphoethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (SPH), or a combination thereof.

3. The in vitro, cell-free placenta model of claim 1 , wherein the model further comprises at least in a portion of the bilayer a single lipid (a unilipid) or a combination of multiple lipids (a multi-lipid).

4. The in vitro, cell-free placenta model of claim 1 , comprising human trophoblast cell line-extracted lipids.

5. The in vitro, cell-free placenta model of claim 1 , comprising human placenta tissue-extracted lipids.

6. The in vitro, cell-free placenta model of claim 1 , further comprising an acceptor compartment configured on the opposite side of the bilayer as the donor compartment, such that the model is capable of simulating the test compound's permeation through an in vivo human placenta by the test compound's permeation from the donor compartment through the in vitro bilayer and into the acceptor compartment.

7. The in vitro, cell-free placenta model of claim 1 , wherein the detector comprises a quartz crystal microbalance operative to provide frequency changes (ΔF) and/or dissipation changes (ΔD), (QCM-D), disposed on a side of the bilayer, such that the model is capable of simulating the test compound's adsorption on or into an in vivo human placenta by a QCM-D monitoring of a viscoelasticity of the in vitro bilayer.

8. The in vitro, cell-free placenta model of claim 1 , further comprising wherein the detector comprises electrodes on both sides of the in vitro bilayer, said electrodes capable of providing a measurement of changes in electrical conductivity after the test compound is introduced into the donor chamber.

9. The in vitro, cell-free placenta model of claim 1 , wherein the detector comprises a UV-Vis absorbance detector, a mass spectrometry detector, a conductivity detector, a colorimetric detector, a fluorescence detector, or a combination thereof.

10. The in vitro, cell-free placenta model of claim 1 , wherein the membrane supporting the lipid bilayer comprises a polymer and/or placental collagen.

Assignments (2)

Continuity (4)

References Cited (346)