Improved methods of cell culture for adoptive cell therapy

A cell culture and cell technology, applied in the field of cell culture, can solve problems such as the complexity of T cell methods

Active Publication Date: 2015-03-11
WILSON WOLF MFG
View PDF7 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this further complicates the method of making T cell

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Improved methods of cell culture for adoptive cell therapy
  • Improved methods of cell culture for adoptive cell therapy
  • Improved methods of cell culture for adoptive cell therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Demonstration of the limitations of conventional methods.

[0086] The data in this example demonstrates that when using 2ml medium volume per well (i.e., medium height is 1.0cm, medium volume to surface area ratio is 1ml / cm 2 ) standard 24-well tissue culture plate (that is, each well has a surface area of ​​2 cm 2 ) in the limitations of conventional culture methods for preparing EBV-CTL.

[0087] Stage 1 of culture, day 0: γ-irradiated (40Gy) antigen-presenting autologous EBV-LCL at a ratio of 40:1 (PBMC:LCL) and 1 ml / cm 2 The ratio of medium volume to growth surface for cell composition from normal donor PBMC (approximately 1×10 6 cells / ml) were cultured to initiate expansion of the EBV-CTL population, whereby 45% Click medium (Irvine Scientific, Santa Ana, CA), 2mM GlutaMAX-I, and 10% FBS supplemented RPMI 1640 built in approximately 1×10 6 cells / cm 2 surface density of the cell composition.

[0088] Stage 2 of culture, days 9-16: On day 9, EBV-CTL...

Embodiment 2

[0100] Example 2: At any given stage or at the beginning of each stage of culture, a reduction in the amount of time required to increase a target cell population can be achieved by reducing the cell surface density of the target cell population.

[0101] We hypothesized that the reduced expansion rate of the target cell population after the second T cell stimulation compared to the first stimulation is due to restrictive cell culture conditions leading to activation-induced cell death (AICD). For example, refer to image 3 A, At first stimulation, the EBV antigen-specific T cell component of PBMCs constituted at most 2% of the population, and thus antigen-specific responsive T cells were seeded at a density of less than 2 × 10 4 / cm 2 , the remaining PBMC acted as non-proliferative feeder cells (shown as image 3 CFSE-positive cells in A), these feeder cells maintain optimal cell-cell contacts that enable the proliferation of antigen-specific CTLs. In contrast, at the seco...

Embodiment 3

[0105] Example 3: Minimum Surface Density of Cell Populations Comprising Target Cells and / or Antigen Presenting Cells can allow growth of target cell populations seeded at very low surface densities.

[0106] Figure 4 shows when continuing image 3 The work described in is an example of the results we have obtained, which further demonstrates that when the target cells need the support of other cells, as long as the target cells are provided with sufficient feeder cells and / or antigen presenting cells, unconventional low The target cell surface density can then initiate population expansion. In these experiments, we went on to demonstrate that with an R:S ratio of about 1.0×10 6 target cells / cm 2 Compared to only about 3900 target cells / cm at an R:S ratio of 1 to 32 2 We stopped testing how the total cell composition at a surface density and R:S ratio between the target cells greatly expanded beyond 50-fold the starting surface density.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Inoculation densityaaaaaaaaaa
Surface densityaaaaaaaaaa
Login to view more

Abstract

Production and use of novel therapeutic cells, called T-Vehicles, in the allogeneic Adoptive Cell Therapy setting allows a wide range of therapeutic benefits to accrue with minimal or no risk of GVHD. T-Vehicles are created from donor T cells that are altered to contain therapeutic attributes that do not include their native antigen receptors and can deliver therapeutic benefits irrelevant of their native antigen specificity. T-Vehicles can possess highly restricted native antigen specificity that renders them unable to recognize antigens present on normal cells and incapable of initiating GVHD, making them ideal transport vehicles to deliver various therapeutic attributes in vivo. In essence, production and use of T-Vehicles is a paradigm shift that opens the door to therapeutic application of T cells in ways not previously contemplated, independent of whether or not there is an HLA match between the donor and the recipient.

Description

[0001] related application [0002] This application is a continuation-in-part of U.S. Patent No. 12 / 963,597, filed December 8, 2010, entitled "IMPROVED METHODS OF CELL CULTURE FOR ADOPTIVE CELL THERAPY" (hereinafter "parent application"), which requires Priority to U.S. Provisional Application No. 61 / 267,761, entitled "IMPROVED METHODS OF CELL CULTURE FOR ADOPTIVE CELL THERAPY," filed December 8, 2009, which is incorporated herein by reference in its entirety. technical field [0003] The present invention relates generally to methods of culturing cells, and more particularly to culturing cells for cell therapy. The present invention also relates to the preparation of T cells with therapeutic properties for Adoptive Cell Therapy. Background technique [0004] Cell culture is a major factor in the cost and complexity of cell therapy. With current methods, the process of culturing cells is time-consuming and costly. Typically, a staged in vitro culture process is used in o...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/0783A61K35/17A61P37/00A61K35/51A61K39/00
CPCA61K39/0011A61K35/51C12N5/0638A61K2039/5158A61K39/39A61K39/12C12N2710/16134C12N2710/16234A61P35/00A61P37/00A61K2039/5156C07K14/7051C07K2319/03A61K35/17
Inventor 约翰·R·威尔逊
Owner WILSON WOLF MFG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap