Method of cryopreservation of stem cell-derived retinal pigment epithelial cells on polymeric substrate

Patent metadata

Field	Value
Publication number	US-2017196221-A1
Application number	US-201715418099-A
Country	US
Kind code	A1
Filing date	Jan 27, 2017
Priority date	Apr 29, 2011
Publication date	Jul 13, 2017
Grant date	—

Abstract

Disclosed herein are methods and compositions for the cryopreservation of stem cells, such as stem-cell derived retinal pigment epithelial cells, that have been seeded onto and cultured on a substrate, such as a polymeric substrate. Such cryopreserved stem cells are useful for cell therapies, such as treatment of ocular damage or disease.

First claim

1 . (canceled)

2 . A substrate suitable for cryopreserving a monolayer of seeded cells, stem cells, stem cell derived cells or combinations thereof, thawing, and subsequent implantation at a target site, the substrate comprising:
a non-porous membrane, wherein the substrate has one or more characteristics selected from (i) a coefficient of thermal expansion of the substrate (ii) a substrate elasticity parameter (iii) a substrate thickness (iv) surface modification, said characteristics enhance viability of the seeded cells and functionality of the substrate.

3 . The substrate of claim 2 , wherein the membrane material is selected to yield a thermal coefficient of expansion of the substrate that has reduced adverse impact on the seeded cells during cryopreservation and subsequent thawing.

4 . The substrate of claim 2 , wherein the substrate has a diameter, length, width, thickness, and formulation of materials that can be adjusted to tailor the thermodynamic properties of the substrate.

5 . The substrate of claim 2 , wherein the substrate comprises an elasticity parameter configured to conform to target anatomy during and after implantation.

6 . The substrate of claim 2 , wherein the substrate comprises an elasticity parameter configured to allow the substrate to maintain its integrity and remain unkinked, untorn, and undamaged during and after implantation.

7 . The substrate of claim 2 , wherein the substrate comprises a thickness is selected to retain substrate functionality during the cryopreservation and subsequent thawing temperature changes.

8 . The substrate of claim 2 , wherein the substrate comprises a permeability that is defined by the substrate thickness.

9 . The substrate of claim 2 , wherein the substrate comprises a permeability configured to permit adequate nutrient passage to the cells and/or adequate passage of cellular waste material away from the substrate.

10 . The substrate of claim 2 , wherein the substrate further comprises a surface modification comprising a coating to enhance adhesion and viability during and after the cryopreservation process.

11 . The substrate of claim 10 , wherein the coating comprises one or more of Matrigel, vitronectin, and retronectin.

12 . The substrate of claim 2 , wherein the substrate comprises a surface modification comprising an addition of materials or chemicals to allow for visualization of the substrate in situ.

13 . The substrate of claim 2 , wherein the monolayer of seeded cells comprises retinal pigment epithelial (RPE) cells.

14 . The substrate of claim 2 , wherein the monolayer of seeded cells comprises unpolarized embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells.

15 . The substrate of claim 2 , wherein the non-porous membrane comprises parylene.

16 . The substrate of claim 2 , wherein the non-porous membrane comprises a biodegradable material.

17 . A parylene substrate configured for implantation into a target site, the parylene substrate comprising:
a non-porous membrane configured for cryopreserving a monolayer of one of seeded cells, stems cells, stem cell derived cells, and combinations thereof,
wherein the non-porous membrane is configured to withstand cryopreservation at a temperature reduction rate of between about 10° C. per 10 seconds to about 10° C. per 120 seconds to an intermediate temperature of −90° C. for a first period of time and a storage temperature for a second period of time, and
wherein the non-porous membrane is configured to withstand thawing and subsequent implantation at a target site.

18 . The substrate of claim 17 , wherein the non-porous membrane has a permeability configured to permit adequate nutrient passage to the cells and/or adequate passage of cellular waste material away from the substrate.

19 . The substrate of claim 17 , wherein the non-porous membrane further comprises a coating to enhance adhesion and viability during and after the cryopreservation process.

20 . The substrate of claim 20 , wherein the coating comprises one or more of Matrigel, vitronectin, and retronectin.

Assignees

• Univ Southern California

Inventors

• Zhu Danhong
• Hinton David
• Ahuja Ashish
• Humayun Mark

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P27/02

  Ophthalmic agents · CPC title

• A61P13/12

  of the kidneys · CPC title