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Methods for cryopreserving and encapsulating cells

A cryopreservation and cell technology, applied in animal cells, embryonic cells, cell encapsulation, etc., can solve problems such as limiting the value of cells

Inactive Publication Date: 2014-03-19
ANTHROGENESIS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are certain disadvantages to cryopreserving cells, including damage to cells during freezing and / or thawing and the need to culture cells after thawing to ensure their proper recovery
These disadvantages limit the value of cryopreserved cells, especially when it is desired to use the cryopreserved cells immediately after thawing or shortly thereafter

Method used

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  • Methods for cryopreserving and encapsulating cells
  • Methods for cryopreserving and encapsulating cells

Examples

Experimental program
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Effect test

Embodiment 1

[0092] 7.1 Example 1: Viability of Encapsulated Placental Stem Cells

[0093] This example demonstrates that placental stem cells are viable when encapsulated in alginate.

[0094] 7.1.1 Store at 25°C and 37°C

[0095] Placental stem cells (5×10 5 cells / ml) were suspended in 0.75% alginate solution and added by dropping the cell / liquid alginate solution into CaCl 2 solution (1.5% w / v) for encapsulation. The encapsulated cells were stored at 25°C or 37°C for seven days, and cell viability was measured at various time points by labeling dead cells with propidium iodide and live cells with acetylmethoxy (AM) calcein.

[0096] The encapsulated placental stem cells were kept viable for 7 days at 37°C and 2 days at 25°C.

[0097] 7.1.2 refrigeration

[0098] Placental stem cells (5×10 5 cells / ml and 2 x 10 6 cells / ml) were suspended in 0.75% alginate solution and added by dropping the cell / liquid alginate solution into CaCl 2 solution (1.5% w / v) for encapsulation. Wil...

Embodiment 2

[0100] 7.2 Example 2: Encapsulation of cryopreserved placental stem cells

[0101] This example demonstrates that cryopreserved placental stem cells can be encapsulated post-thaw and they remain viable without the need to culture the post-thaw cells.

[0102] 7.2.1 Add alginate after thawing and encapsulate after thawing

[0103] Placental stem cells (7.5 × 10 6 cells / ml) were thawed and mixed 1:1 with 1.5% alginate (in PBS) without any intermediate culture steps. Cells were then encapsulated as described in Example 1. After encapsulation and without any intermediate culture steps, the (BioLife Solutions, Bothell, WA) storage solution was added to the encapsulated placental stem cells, and the cells were stored in syringes at 4°C for 7 days (i.e. the cells were stored at 100% middle). According to the manufacturer's instructions, use The viability of encapsulated placental stem cells was analyzed with a cell viability analyzer (Beckman Coulter, Brea, CA). Immedia...

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Abstract

Described herein are methods of cryopreserving cells that have been suspended in alginate as well as methods for encapsulating cryopreserved cells that have been suspended in alginate. Further provided herein are cellular compositions comprising cells that have been processed in accordance with the methods described herein.

Description

[0001] This application claims priority to US Provisional Patent Application No. 61 / 428,427, filed December 30, 2010, the disclosure of which is incorporated herein by reference in its entirety. 1. Technical field [0002] Described herein are methods of cryopreserving cells that have been suspended in alginate and methods of encapsulating cryopreserved cells that have been suspended in alginate. Also provided herein are cellular compositions comprising cells that have been treated according to the methods described herein. 2. Background technology [0003] Cryopreservation is a method in which cells can be preserved by cooling them to low temperatures. At these low temperatures, biological activity is effectively stopped, including biochemical reactions that would normally lead to cell death. Thus, cryopreservation provides a valuable method for storing cells for later use. However, there are certain disadvantages to cryopreserving cells, including damage to cells during ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K9/14A61K35/50
CPCA61K9/14C12N5/0012A61K35/50A61K35/12C12N5/0605A01N1/0231A61P43/00A01N1/0284
Inventor 萨沙·阿布拉门逊
Owner ANTHROGENESIS CORP
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