Methods of manufacture of therapeutic products comprising vitalized placental dispersions

a technology of vitalized placental dispersions and placental products, which is applied in the direction of biocide, drug composition, peptide/protein ingredients, etc., can solve the problems of high morbidity and mortality rate, and insufficient treatment, and achieves the effect of limited digestion

Inactive Publication Date: 2011-09-01
OSIRIS THERAPEUTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, burns produce local tissue damage as well as systemic consequences.
Burn wounds continue to be a frustrating and serious problem in the clinic, and these wounds are often accompanied by high morbidity and mortality rates.
However, for severe and large surface area burns, this treatment is not satisfactory.
However, the amount of skin available for grafting is often extremely limited, and this procedure always results in donor site wounds.
The effectiveness of placental membranes such as amniotic membranes for burns was recorded in a number of published reports; however, the use of placental membranes for large surface area burns is limited due to challenges in providing sufficient placental membranes to cover large areas.

Method used

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  • Methods of manufacture of therapeutic products comprising vitalized placental dispersions
  • Methods of manufacture of therapeutic products comprising vitalized placental dispersions
  • Methods of manufacture of therapeutic products comprising vitalized placental dispersions

Examples

Experimental program
Comparison scheme
Effect test

example 1

Obtaining Placental Tissue

[0176]A whole placenta was obtained from a registered tissue bank after informed consent. The placenta and placed, with the maternal surface (rough surface) face down, on a sterile tray. The amniotic-chorionic membrane was cut and removed from the placenta. The chorionic membrane was then separated from the amnion and washed twice in PBS.

[0177]The chorionic membrane was then soaked in an anticoagulant (ACD-A) solution to remove blood clots and then washed again in PBS.

[0178]The chorionic membrane was then digested by incubation with dispase II for 30 min. at 37° C. The trophoblast layer was mechanically removed by scraping with fingers and the chorion was washed again in PBS.

[0179]The chorionic membrane was then incubated for 24 hours in an antibiotic cocktail containing gentamicin, vancomycin, and amphotericin B, and washed again in PBS.

example 2

Digesting Placental Tissue

[0180]A chorion membrane (obtained from Example 1) was digested by incubation in 200 mL of a collagenase II solution (300 U / mL in DMEM) for 10 min at 37° C. The chorionic membrane was then removed, leaving a digestion suspension containing collagenase and placental cells.

[0181]The volume and container for digestion was determined based on the need to provide a suitable digestion environment for the tissue once placed on a shaker. The digestion was carried out on a standard plate shaker set at moderate speed in a 37° C. cell culture incubator.

example 3

Obtaining Placental Cells

[0182]A digestion suspension comprising placental cells (obtained from Example 2) was centrifuged at 2000 rcf for 5 min to separate the digestive enzyme (collagenase II) from the placental cells. This step centrifugation step may enhance cell viability by preventing over-digestion and ensure that the enzyme is washed away before homogenizing the tissue. This centrifugation step pellets the cells without damaging them, allowing the collagenase II to be removed as supernatant.

[0183]The cells were then centrifuged again, the supernatant poured off, and the placental cells were resuspended in a small volume (2 mL) of cryprotectant (5% DMSO in saline). Two mL provides an adequate volume to resuspend the cells while not over-diluting the chorion membrane dispersion once the cells have been added.

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Abstract

This invention provides a fluid therapeutic placental product comprising placental cells and a placental dispersion comprising placental factors. The placental cells and the placental dispersion are derived from placental tissue. A placental tissue can optionally be an amnion, chorion, or a trophoblast-depleted chorion. The placental product of the present invention is useful in treating a patient with a tissue injury (e.g. wound or burn) by applying the placental product to the injury. Similar application is useful with ligament and tendon repair and for engraftment procedures such as bone engraftment.

Description

RELATED APPLICATIONS[0001]This application claims priority to:[0002]U.S. Provisional Application Ser. No. 61 / 338,464 entitled “Selectively Immunodepleted Chorionic Membranes”, filed on Feb. 18, 2010 bearing Docket No. 22924US01,[0003]U.S. Provisional Application Ser. No. 61 / 338,489 entitled “Selectively Immunodepleted Amniotic Membranes”, filed on Feb. 18, 2010 bearing Docket No. 22925US01, and[0004]U.S. Provisional Application Ser. No. 61 / 369,562 entitled “Therapeutic Products Comprising Vitalized Placental Dispersions filed on Jul. 30, 2010 bearing Docket No 23498US01, the contents of which are hereby incorporated by reference in their entireties.[0005]This application is being co-filed on Feb. 18, 2011 with, and incorporates by reference, applications entitled:[0006]“Immunocompatible Chorionic Membrane Products”,[0007]“Methods of Manufacture of Immunocompatible Chorionic Membrane” Products,[0008]“Immunocompatible Amniotic Membrane Products”,[0009]“Methods of Manufacture of Immuno...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/50A61P17/02
CPCA01N1/0221A61K35/50A61K38/1825A61K38/1841C12N2501/115A61K38/57A61K35/28C12N5/0605A61K38/39C12N2500/02C12N2502/025A61P17/00A61P17/02A61P43/00
Inventor JANSEN, TIMOTHYTOM, SAMSONDANILKOVITCH, ALLAYOO, DANAZERHUSEN, JAIME
Owner OSIRIS THERAPEUTICS
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