Shrimp Sperm Cryopreservation via Partial Homogenization

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current cryopreservation protocols for shrimp spermatophores and sperm mass often result in the loss of original physical properties and fertilizing capacity, as they require complete disintegration and homogenization of samples for effective preservation, which is not suitable for maintaining intact viability and morphology.

Innovation Solution

A protocol involving an initial extender solution with specific concentrations of NaCl, KCl, H3BO3, MgSO4, and antibiotics, followed by Trehalose treatment and partial submersion in a cryoprotectant solution containing DMSO, allowing for cryopreservation without complete disintegration, thereby preserving the original physical and fertilizing properties of shrimp sperm samples.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If complete disintegration and homogenization of samples is performed for cryopreservation, then effective preservation is achieved, but original physical properties and fertilizing capacity are lost

Engineering Contradiction:
Improvepreservation effectivenessVSAvoidoriginal physical properties
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent applies partial action by performing only the necessary degree of homogenization to enable cryoprotectant penetration, rather than complete disintegration. The sperm packet is partially homogenized to achieve adequate mixing with cryoprotectants while deliberately maintaining the integrity of the sperm packet structure, thus preserving original physical properties and fertilizing capacity.

Inventive Principle:
Principle #16Partial or excessive action

2Ease of operation

If complete disintegration of samples is performed, then cryoprotectant contact is improved, but sperm packet integrity is lost

Engineering Contradiction:
Improvecryoprotectant contactVSAvoidsperm packet integrity
Core Design Contradiction:
Ease of operationVSShape

Solution Approach 1:

The method performs partial homogenization - enough to ensure adequate contact between cryoprotectants and sperm cells, but not enough to destroy the sperm packet structure. This intermediate level of processing achieves the necessary cryoprotectant penetration while maintaining the structural integrity required for successful fertilization.

Inventive Principle:
Principle #16Partial or excessive action

3Reliability

If DMSO is used as cryoprotectant agent, then cryopreservation is achieved, but osmotic balance requires additional compounds

Engineering Contradiction:
Improvecryopreservation effectivenessVSAvoidsolution composition
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent uses a composite cryoprotectant solution containing DMSO combined with other compounds that maintain osmotic balance. This composite approach leverages the cryoprotective properties of DMSO while supplementing it with additional substances that ensure proper osmotic conditions, achieving effective cryopreservation through material composition rather than procedural complexity.

Inventive Principle:
Principle #40Composite materials

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The protocol effectively maintains the viability and morphological integrity of shrimp sperm samples, enabling successful fertilization and preserving the original consistency of the samples, with treatment B (Trehalose 0.25-0.5 M and DMSO 10-15%) demonstrating 100% intact cell membrane and viability post-thawing.

Implementation Method 1

this is achieved through the addition of cryoprotectant substances that prevent crystal formation during the freezing process either from reducing the quantity of intracellular water and/or from altering their behavior by modifying the attractive forces between the water molecules, for example by breaking the hydrogen bonds

Methodology Applied
Scientific EffectCryoprotection:

Implementation Method 2

modifying the attractive forces between the water molecules, for example by breaking the hydrogen bonds

Methodology Applied
Scientific EffectHydrogen bonding:

Implementation Method 3

A protocol involving an initial extender solution with specific concentrations of NaCl, KCl, H3BO3, MgSO4, and antibiotics, followed by Trehalose treatment and partial submersion in a cryoprotectant solution containing DMSO

Methodology Applied
Scientific EffectVitrification: Vitrification

Implementation Method 4

The protocol effectively maintains the viability and morphological integrity of shrimp sperm samples, enabling successful fertilization and preserving the original consistency of the samples

Methodology Applied
Scientific EffectCryopreservation: Cryogenics

Data Source

PatentEP3516959B1Protocol for the cryopreservation of shrimp spermatophores/sperm mass
Publication Date: 2021.11.10 CENT DE INVESTIGACION CIENTIFICA Y DE EDUCACION SUPERIOR DE ENSENADA BAJA CALIFORNIA CICESE
  • EP3516959B1 patent drawingFigure 1
  • EP3516959B1 patent drawingFigure 2
  • EP3516959B1 patent drawingFigure 3a~3b

AI summary

A protocol for the cryopreservation of high viscosity biological samples, preferably of spermatic samples of crustaceans or insects, that enables maintaining intact the physical properties of the original sample preserving the viability of the sperm packet.