Unlock instant, AI-driven research and patent intelligence for your innovation.

Cryopreservation of cells

A cell and plant cell technology, applied in the field of cryopreservation of transformed and non-transformed cells, can solve the problem of providing less time for cryopreservation

Inactive Publication Date: 2007-10-03
DOW AGROSCIENCES LLC
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Little information is available in the prior art on the length of refrigeration (usually measured in months or even hours), and there is limited information on whether cells are able to grow indefinitely or at least to the desired number of passages under common culture conditions
In addition, there are few data revealing the genetic and production effects of target genes or gene products (both from original pluripotent seed stocks and expanded and refrigerated working seed stocks) after long-term storage or after long-term culture after removal from storage. stability

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
  • Cryopreservation of cells
  • Cryopreservation of cells
  • Cryopreservation of cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Refrigeration

[0066] The media and components used in this example are listed in Tables 2-4. Components supplied by suppliers are also listed. Cells to be refrigerated are grown in shake flasks containing NT1 medium at 25°C; split cells 1:3 (or 30% seeded) during the mid-log (mid-exponential) growth phase (3-4 days after seeding the flask) , Passage for at least 1-10 generations (one embodiment is at least 6 generations). Clean the outer surface of the culture flask with 1% sodium perchlorate solution before transferring to a sterile biological safety cabinet, and wipe the outer surface of the culture flask with a sterile alcohol pad before transferring the cells to a sterile 225ml centrifuge tube. The cells were centrifuged at 1000 RPM for 1 min at 4°C, and the supernatant was removed with a sterile pipette. Resuspend the cells at the original volume with the appropriate medium and transfer to a sterile 1-liter Erlenmeyer flask, add an equal volume of r...

Embodiment 2

[0069] Example 2 - Thawing of Refrigerated Tobacco Cells

[0070] Single tubes of cells or collections of multi-tube cells can be thawed. Remove the tubes from the storage unit and place on dry ice. Submerge it in a 45°C water bath, gently moving the tube racks in the water bath to help promote rapid and consistent thawing of the tubes. After approximately 2.5 minutes (to the point of just thawing), gently invert the tube 3 times to mix the cells that have settled to the bottom of the tube. In a laminar flow cabinet, aspirate 2 ml of cells from a collection of multiple tubes or a single tube onto 8-10 layers of sterile 70 mm No. 4 Whatman filter paper in a sterile Petri dish, cover and allow to drain for 2 minutes.

[0071] At this point a small number of cells (~0.5ml) were stained for TTC viability. After draining for 2 minutes, the top filter paper with cells was transferred to semi-solid NTB1 medium without bialaphos selection. Plates were then wrapped with 3M tape and...

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

No PUM Login to View More

Abstract

The present invention relates to methods for the cryopreservation of transformed and non-transformed cells. Also provided by the subject invention are methods of recovering cells that have been cryopreserved. Cultures of cells that have been successfully recovered from cryopreservation are also provided.

Description

field of invention [0001] This application claims priority to US Provisional Application Serial No. 60 / 625,401, filed November 5,2004. field of invention [0002] The present invention relates to methods of cryopreservation of transformed and non-transformed cells. Also provided by the present invention are methods of recovery of cryopreserved cells. And provide cell cultures that have successfully recovered from cold storage. Background of the invention [0003] The "universal seed" principle for the production of biopharmaceuticals and bioagrochemicals utilizes living organisms as part of the manufacturing process and is based on some fundamental principles: 1) Monocultures of defined origin and passage history are deposited that have Cell phenotype with defined characteristics and desired manufacturing characteristics; 2) long-term (spanning several years or more) preservation (especially cold storage); 3) cells can be recovered, expanded, passaged indefinitely as "wo...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/02C12N5/04C12N5/14
CPCA01N3/00A01N1/02A01N1/0278C12N5/04C12N5/14C12M1/00
Inventor W·M·爱因雷J·R·博林格尔C·M·L·拉尔森M·卢L·Y·沈P·S·亚佳库马尔R·J·佳里森D·R·帕雷迪
Owner DOW AGROSCIENCES LLC