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

Compositions for cryopreservation and methods of use thereof

A technology for compositions and compounds, applied in biochemical equipment and methods, drug combinations, preservation of human or animal bodies, etc., which can solve problems such as cell death, cell damage, external plasma membrane and internal organelle puncture, etc.

Pending Publication Date: 2020-09-11
常青树生物科学公司
View PDF14 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cryopreservation itself, and the freeze-thaw cycles associated with cryopreservation, damage cells by forming ice crystals in close proximity (and sometimes intracellularly), again causing puncture and rupture of the outer plasma membrane and inner organelles (so-called cryopreservation). melting injury)
The combination of isolation and cell damage associated with cryopreservation can also lead to transdifferentiation of cells from one differentiated state to another cell type, or dedifferentiation of cells, thereby further complicating the subsequent use of any isolated cells
[0007] Current approaches do not address the subsequent cell damage and death due to these damages and leave cells and tissues and organs overly damaged
Furthermore, no solution has been implemented to prevent subsequent necrosis
Likewise, little, if any, progress has led to improvements in cryopreservation techniques, and it is unclear what molecular pathways exactly lead to cell death during freeze-thaw

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
  • Compositions for cryopreservation and methods of use thereof
  • Compositions for cryopreservation and methods of use thereof
  • Compositions for cryopreservation and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0201] Example 1. Cryopreservation solutions reduce the levels of reactive oxygen species (ROS), apoptosis, and necrosis resulting from tissue dissection and cell manipulation.

[0202] Histological anatomy and cellular manipulation of mouse cortical neuronal tissues were studied in the presence and absence of cryopreservation solutions (Soln, soln). (for Figure 1A-1C Soln / soln: 20nM 5-((7-chloro-1H-indol-3-yl)methyl)-3-methyl-2,4-imidazolidinedione), 5nM 3, 6-dibromo-α-(1-piperazinylmethyl)-9H-carbazole-9-ethanol dihydrochloride, 0.05mM nicotinamide adenine dinucleotide (NAD), 0.01μM adenosine triphosphate (ATP) , 0.01 μM cyclosporine A, and 0.1 Kuntz units manganese superoxide dismutase. ) tissue dissection and cell manipulation of freshly dissected tissues (fresh) produced reactive oxygen species (ROS) (Fig. 1A), whereas in non-dissected tissues (mock), almost no ROS were observed. Freshly dissected tissue (fresh) produced several-fold increased ROS generation, while incl...

example 2

[0204] Example 2. Cryopreservation solutions reduce levels of reactive oxygen species (ROS), apoptosis, and necrosis resulting from freezing and subsequent thawing of cells.

[0205] Cell freezing / thawing of neurons was studied for levels of ROS, apoptosis and necrosis markers over time in a number of cell types. (For Figure 2A-2C Soln: 20nM 5-((7-chloro-1H-indol-3-yl)methyl)-3-methyl-2,4-imidazolidinedione), 5nM 3,6- Dibromo-α-(1-piperazinylmethyl)-9H-carbazole-9-ethanol dihydrochloride, 0.05mM nicotinamide adenine dinucleotide (NAD), 0.01μM adenosine triphosphate (ATP), 0.01 μM cyclosporine A, and 0.1 Kuntz units manganese superoxide dismutase. ) (CypD-cyclophilin D) Freezing / thawing of neurons generates ROS (Nox1 detected) (frozen), which is almost completely abolished by including cryopreservation solution (+Soln) compared to tissue before freezing (mock) The effect (Fig. 2A). Apoptosis was slightly enhanced after freezing / thawing (freezing) compared to tissue before fr...

example 3

[0206] Example 3. Motor neuron stem cells frozen in cryoprotectant solution maintain motor neuron identity.

[0207] Cell freeze / thaw cycles cause stem cells to lose their identity, as measured by CHAT1 levels in motor neurons. Cultured and expressed CHAT1 (fresh) when compared to stem cells frozen in the absence of these compounds, and then in a solution containing 0.01 μM cyspA (cyclosporine A), or 20 nM Necl, or 5 nM iMAC2 Frozen stem cells (motor neuron precursors) maintained CHAT1 levels, showing that cells after freezing (frozen) had very low CHAT1+ levels ( image 3 ). as shown here ( image 3 ), each of cyspA, Nec1 and IMAC2 provides cryoprotection to stem cells after freezing, maintaining cell identity. Cryoprotectants are used to freeze cells in solution, maintaining motor neuron identity, as measured by levels of CHAT1.

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

Provided herein are compositions for cryopreservation of mammalian cells, tissues, and organs. The compositions include a necroptosis inhibitor compound and a Bax channel inhibitor compound. Providedherein are also methods of use of the compositions, for treating, preventing, inhibiting, or reducing the incidence of cellular plasticity, or necroptosis or necrosis, of a plurality of cells, whereinthe cells are brought into contact with the composition.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application 62 / 612,552, filed December 31, 2017, which is hereby incorporated by reference in its entirety. technical field [0003] Compositions and methods thereof for cryopreservation of mammalian cells, tissues and organs are described herein. Background technique [0004] Excessive cell death is a major obstacle to the clinical use of cells, tissues and organs obtained from living donors or grown in culture. This problem is especially acute when there is a protracted period of time between obtaining the cells and using them, requiring stabilization of the cells during prolonged storage. In general, the process of isolating donor cells, tissues, or organs itself damages the cells as it involves enzymatic or mechanical removal, enzymatic digestion of extracellular proteins, transfer of cells between vessels, centrifugation, resuspension in solution, and fil...

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): A61K31/415A61K31/4155A61K31/4178
CPCA61K31/4166A61P9/10C12Y115/01001A61P1/18A61K38/446A61K38/1761A61P25/28A61K31/415A61K31/52A61K31/4178A61K31/495A61K2300/00A61K47/549A61P43/00A01N1/0226A61K31/7084A61K31/496A61K38/13A01N1/0221A61K31/7076
Inventor J·迪登贝格P·伊塞罗维奇
Owner 常青树生物科学公司