Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells

A technology of human placenta and perfusate, applied in blood/immune system cells, antitumor drugs, animal cells, etc., can solve the problems of difficult to maintain tumor targeting, kill tumors, and difficult to handle.

Active Publication Date: 2013-10-23
ANTHROGENESIS LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Despite the favorable properties of NK cells in killing tumor cells and virus-infected cells, handling them and using them for immunotherapy remains difficult, mainly due to the difficulty in maintaining their tumor targeting during culture and expansion. and the ability to kill tumors

Method used

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  • Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells
  • Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells
  • Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells

Examples

Experimental program
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Effect test

Embodiment 2

[0377] 6.2. Example 2: Placental intermediate natural killer from mixed placental perfusate and cord blood Cell characterization

[0378] Mix the donor-matched monocytes (mixed) of umbilical cord blood and placental lavage fluid, and wash once with FACS buffer (RPMI-1640 containing 5% FBS), and use the antibodies listed in Table 2, in BDFACSCanto (BD Biosciences) for immunophenotyping. Use FlowJo software (Tree Star) to analyze the data.

[0379] Table 2: List of antibodies used in immunophenotyping

[0380] Item

Sellers

Catalog number

FITC anti-human CD3

BD Bioscience

555332

FITC anti-human CD3

Miltenyi

130-080-401

APC-Cy7 anti-human CD3

BD Bioscience

557832

FITC anti-human CD16

BD Bioscience

555406

PE-Cy5 anti-human CD16

BD Bioscience

555408

PE anti-human CD56

BD Bioscience

555516

PE anti-human CD56

Miltenyi

130-090-755

PE-CY5 anti-human CD56

BD Bioscience

555517

PE-Cy7 anti-human CD56

BD Bioscience

557747

PE anti-human CD94

R&D

FAB-1058P

...

Embodiment 3

[0410] 6.3. Example 3: Cytotoxicity of natural killer cells to tumor cells

[0411] This example demonstrates that placental intermediate natural killer cells are cytotoxic to tumor cells. As confirmed in the cytotoxicity test and NK cell cytokine secretion Luminex analysis, PINK cells from HPP are cytotoxic to acute myeloid leukemia cells.

[0412] In the cytokine secretion test, NK cells enriched with CD56 microbeads from HPP were mixed with KG-1a acute myeloid leukemia cells at a ratio of 1:1. After 24 hours of culture, the supernatant was collected and subjected to Luminex analysis of IFN-γ and GM-CSF secretion. As shown in Figure 2, after co-culturing CD56-enriched HPP cells with KG-1a cells for 24 hours, increased levels of IFN-γ and GM-CSF were observed.

[0413] Cytotoxicity of PINK cells

[0414] In the cytotoxicity test using PINK cells, target tumor cells were labeled with carboxyfluorescein succinimidyl ester (CFSE). CFSE is a vital dye that is non-toxic to cells, and ...

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Abstract

Provided herein are placental perfusate, placental perfusate cells, and placenta-derived intermediate natural killer (PINK) cells, and combinations thereof. Also provided herein are compositions comprising the same, and methods of using placental perfusate, placental perfusate cells, and placenta-derived intermediate natural killer cells, and combinations thereof, to suppress the growth or proliferation of tumor cells, cancer cells, and the like, and to treat individuals having tumor cells. In a preferred embodiment, the composition comprises: solated CD56+, CD16- natural killer cells.

Description

[0001] This application is a division of the invention application with the application date of September 29, 2008, the application number of 200880118399.6, and the title "Tumor suppression using human placental lavage fluid and human placental intermediate natural killer cells". [0002] This application claims the right of preference for the U.S. Provisional Patent Application No. 60 / 995,763 filed on September 28, 2008 and the U.S. Provisional Patent Application No. 61 / 090,555 filed on August 21, 2008, the entire contents of which are incorporated by reference Include this article. 1. Field of Invention [0003] The present invention provides by combining tumor cells with placental perfusate, cells from placenta, natural killer cells from placenta (e.g. from placental perfusate), and / or natural killer cells from placenta (e.g. from placental perfusate). A method in which killer cells are contacted with mixed natural killer cells derived from umbilical cord blood to inhibit the g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/50A61P35/00A61K35/12C12N5/0783
CPCA61K2035/124C12N5/0646A61K35/50A61P35/00A61K2239/49A61K39/4613A61K2239/48A61K2239/52A61K39/4644A61K35/17A61K45/06
Inventor 晓奎·张瓦内萨·A·沃斯基纳里安-贝尔斯琳·康尼拉弗·迪利普·帕德里亚
Owner ANTHROGENESIS LLC
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