Nitya G Ray

US Patent:

20020123809, Sep 5, 2002

Filed:

Dec 26, 2001

Appl. No.:

10/032929

Inventors:

Hsin-Chien Tai - Hackensack NJ, US
Alla Laufer - Brooklyn NY, US
Ying Song - Franklin Park NJ, US
Nitya Ray - East Hanover NJ, US

International Classification:

A61F002/02

US Classification:

623/023720, 435/374000

Abstract:

Processes are described for making a cryopreserved Composite Living Construct (CCLC) as well as a corresponding thawed and rinsed CCLC, comprised of separated layers of cultured fibroblasts and cultured keratinocytes, wherein the percent of cells that are viable, i.e., the cell viability, of such CCLC is at least about70 %. The viable cell density in the CCLC is at least about 50% of that before cryopreservation. The storage stability of the CCLC is at least about 12 months. Additionally, the metabolic activity of thawed and rinsed CCLC is at least about 50% of the Composite Living Construct (CLC) before cryopreservation. The structural integrity of CCLC is substantially the same as the CLC before cryopreservation. The process for making the CCLC comprises the steps of: providing a collagen substrate comprised of a collagen sponge layer and a nonporous to cells, semipermeable collagen layer; seeding and culturing, in the presence of a cell growth medium, fibroblasts on and within the collagen sponge layer and keratinocytes on the nonporous to cells, semipermeable collagen layer, thereby providing a CLC; equilibrating the CLC, according to a defined equilibration program with a cryoprotectant solution comprising at least chondroitin sulfate and dimethylsulfoxide; lowering the temperature, according to a programmed rate, to about -90 C.; and storing the CCLC at about -150 C. or lower. The process for preparing the CCLC to treat wounds in humans and in animals additionally comprises programmed thawing as well as a rinsing sequence to substantially remove the cryoprotectants.

US Patent:

20030077816, Apr 24, 2003

Filed:

Dec 26, 2001

Appl. No.:

10/032925

Inventors:

Richard Kronenthal - Rutherford NJ, US
Nitya Ray - East Hanover NJ, US
Kimberly Tahan - Kinnelon NJ, US
John Wilson - New Brighton MN, US

International Classification:

C12M003/04

US Classification:

435/297200, 435/299100, 435/177000, 435/402000

Abstract:

A bioreactor is described comprising: at least one cartridge having disposed therein at least one collagen substrate comprised of a collagen sponge layer and a nonporous to cells, semipermeable collagen layer; a substrate support that retains the collagen substrate within the cartridge; a first compartment between the collagen sponge layer and the inner surface of the first side of the cartridge casing and a second compartment between the nonporous to cells, semipermeable collagen layer and the inner surface of the second side of the cartridge casing; inlet and outlet means for transferring a first medium and a first cell type to the first compartment and inlet and outlet means for transferring a second medium and a second cell type to the second compartment. Also described is a method for using the bioreactor to grow a Composite Living Construct (CLC) comprised of a first layer comprising a cultured first cell type and a second layer comprising a cultured second cell type; seeding and culturing the first cell type on and within the collagen sponge layer; and seeding and culturing the second cell type on the nonporous to cells, semipermeable collagen layer. Another embodiment of this invention includes methods for cutting the CLC into sections or units in preparation for product packaging of appropriate sizes. Yet another embodiment of this invention includes equilibration of the CLC with cryoprotectant solutions within the cartridge, further cutting the CLC into sections or units in preparation for cryopreservation. Yet another embodiment of this invention includes equilibration of the CLC with cryoprotectant solutions and cryopreservation within the cartridge, with the cartridge providing product packaging. Yet another embodiment of this invention includes methods for cutting the CLC into sections or units in preparation for product packaging, followed by equilibration with cryoprotectant solutions and cryopreservation within the package.

US Patent:

20040053409, Mar 18, 2004

Filed:

Aug 15, 2003

Appl. No.:

10/641655

Inventors:

Hsin-Chien Tai - Hackensack NJ, US
Alla Laufer - Brooklyn NY, US
Ying Song - Franklin Park NJ, US
Nitya Ray - East Hanover NJ, US

Assignee:

Ortec International, Inc. - New York NY

International Classification:

C12N005/08

US Classification:

435/374000, 435/397000

Abstract:

Processes are described for making a cryopreserved Composite Living Construct (CCLC) as well as a corresponding thawed and rinsed CCLC, comprised of separated layers of cultured fibroblasts and cultured keratinocytes, wherein the percent of cells that are viable, i.e., the cell viability, of such CCLC is at least about 70%. The viable cell density in the CCLC is at least about 50% of that before cryopreservation. The storage stability of the CCLC is at least about 12 months. Additionally, the metabolic activity of thawed and rinsed CCLC is at least about 50% of the Composite Living Construct (CLC) before cryopreservation. The structural integrity of CCLC is substantially the same as the CLC before cryopreservation. The process for making the CCLC comprises the steps of: providing a collagen substrate comprised of a collagen sponge layer and a nonporous to cells, semipermeable collagen layer; seeding and culturing, in the presence of a cell growth medium, fibroblasts on and within the collagen sponge layer and keratinocytes on the nonporous to cells, semipermeable collagen layer, thereby providing a CLC; equilibrating the CLC, according to a defined equilibration program with a cryoprotectant solution comprising at least chondroitin sulfate and dimethylsulfoxide; lowering the temperature, according to a programmed rate, to about -90 C.; and storing the CCLC at about -150 C. or lower. The process for preparing the CCLC to treat wounds in humans and in animals additionally comprises programmed thawing as well as a rinsing sequence to substantially remove the cryoprotectants.