Systems and methods for in vitro and in vivo imaging of cells on a substrate

Patent metadata

Field	Value
Publication number	US-9274095-B2
Application number	US-201514791007-A
Country	US
Kind code	B2
Filing date	Jul 2, 2015
Priority date	Jul 12, 2010
Publication date	Mar 1, 2016
Grant date	Mar 1, 2016

Abstract

Disclosed herein are generally to methods and systems that facilitate imaging of cells on a substrate and more particularly to pre-implantation (in vitro) and post-implantation (in vivo) imaging of cell-seeded substrates implanted in target tissues in the context of stem cell therapy.

First claim

What is claimed is:

1. A computer-implemented method of determining in vitro whether a substrate comprising stem cell-derived retinal pigment epithelium (RPE) cells is acceptable for implantation in an eye, the computer-implemented method comprising:
directing by a light source light at the substrate;
obtaining by a detector an image of the substrate;
determining by a computer system a number of stem cell-derived RPE cells on the substrate;
determining by the computer system whether the number of stem cell-derived RPE cells is at or above a first pre-stored threshold value;
detecting by a detector light transmitted through the substrate;
determining by the computer system a light transmittance level;
determining by the computer system whether the light transmittance level is at or above a second pre-stored threshold value, wherein the determined light transmittance level indicates pigmentation of the stem cell-derived RPE cells, wherein a certain level of pigmentation of the stem cell-derived RPE cells is an indication of cell functionality sufficient for implantation to the eye; and
generating by the computer system a notification that the substrate is acceptable for implantation to the eye based on the determining of whether the number of stem cell-derived RPE cells is at or above the first threshold value and the determining of whether the light transmittance level is at or above the second threshold value,
wherein the computer system comprises a computer processor and an electronic storage medium.

2. The computer-implemented method of claim 1 , wherein the method is repeated over a period of time when the number of stem cell-derived RPE cells is below the first threshold value or the light transmittance level is below the second threshold value.

3. The computer-implemented method of claim 1 further comprising:
determining by the computer system an area of confluence between an outer boundary of the substrate and an outer boundary of stem cell-derived RPE cell growth on the substrate; and
determining by the computer system whether the area of confluence is at or above a third pre-stored threshold value,
wherein the computer system generates the notification that the substrate is acceptable for implantation only when the area of confluence is at or above the third threshold value.

4. The computer-implemented method of claim 1 further comprising:
calculating by the computer system a density of stem cell-derived RPE cells on the substrate; and
determining by the computer system whether the density of stem cell-derived RPE cells is at or above a fourth pre-stored threshold value,
wherein the computer system generates the notification that the substrate is acceptable for implantation only when the density of cells is at or above the fourth threshold value.

5. The computer-implemented method of claim 1 , wherein the substrate further comprises markings configured to assist counting a number of stem cell-derived RPE cells.

6. The computer-implemented method of claim 1 , wherein the computer system comprises one or more computer systems.

7. The computer-implemented method of claim 1 , wherein the determining of the light transmittance level further comprises detecting a level of ambient light and adjusting the determined light transmittance level based on the level of ambient light.

8. The computer-implemented method of claim 1 , wherein the determining of the light transmittance level further comprises blocking the detector from detecting ambient light by covering the detector with a shield apparatus.

9. The computer-implemented method of claim 1 , wherein RPE cells comprise hESC-RPE cells.

Assignees

• Univ Southern California
• Doheny Eye Inst

Inventors

• Humayun Mark
• Ahuja Ashish
• Le Pere Charles

Classifications

• A61F9/0017

  implantable in, or in contact with, the eye, e.g. ocular inserts · CPC title

• G01N21/6486

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title

• A61B3/12

  for looking at the eye fundus, e.g. ophthalmoscopes (A61B3/13 takes precedence) · CPC title

• G06T2207/10004

  Still image; Photographic image · CPC title

• G01N2201/12

  Circuits of general importance; Signal processing · CPC title