CN109769799A

CN109769799A - A kind of human fat tissue saves liquid and preparation method thereof

Info

Publication number: CN109769799A
Application number: CN201910216220.0A
Authority: CN
Inventors: 焦阳; 王铁; 朱迪·德瓦库玛; 杨丽华; 李靖
Original assignee: JIANGSU RE-STEM BIOTECHNOLOGY CO LTD
Current assignee: JIANGSU RE-STEM BIOTECHNOLOGY CO LTD
Priority date: 2019-03-20
Filing date: 2019-03-20
Publication date: 2019-05-21

Abstract

A kind of human fat tissue saves liquid and preparation method thereof.The invention discloses a kind of human fat tissues to save liquid, and the fat of every 1000ml saves liquid -40:50-70g containing D-40, NaCl50-60mmol, KCl20-30mmol, MgSO₄4-8mmol, sucrose 40-60mmol, penicillin G sodium salt: 0.1~0.2g, amphotericin B: 0.1~0.2g, 0.07~0.13g of gentamicin, reduced glutathione 3-5mmol, adenosine 4-8mmol, surplus are deionized water, and the pH value of the preservation liquid is 7.33-7.53.It by each ingredient mixed dissolution in ionized water, dispenses and seals, saved in 0-4 DEG C.Adipose tissue is saved using this preservation liquid, to fat cell death rate is slowed down, mescenchymal stem cell activity in fat is maintained to have remarkable result, in addition, can effectively inhibit bacterium, is conducive to carry out fat further clinical and research application.

Description

A kind of human fat tissue saves liquid and preparation method thereof

Technical field

The present invention relates to liquid field is saved, liquid and preparation method thereof is saved more particularly to a kind of human fat tissue.

Background technique

With the development of medicine, the effect of adipose tissue is also constantly found.Other than storing energy, adipose tissue also has There is active endocrine function, a variety of hormones can be secreted, and participates in adjusting the level of hormone, the biology of mescenchymal stem cell Effect, also constantly scientific research be clinically found that there are huge application prospects.In addition, in shaping medical technology, rouge Fat tissue is also found to have good tissue substitute function, has good market application prospect.Therefore, to adipose tissue Techniques of preserving is with regard to particularly important.

It there is no specific preparation for saving fatty group at present, the preservation liquid of extemporaneous preparation, it is extensive permanent to tend not to Preservation adipose tissue, and Costco Wholesale is higher, and inhibits antibacterial effect undesirable, is unfavorable for the preservation to fat.

Summary of the invention

The invention mainly solves the technical problem of providing a kind of human fat tissues to save liquid, and raw material includes:

D-40-40:50-70g

NaCl:50-60mmol

KCl:20-30mmol

MgSO4:4-8mmol

Sucrose: 40-60mmol

Penicillin G sodium salt: 0.1~0.2g

Amphotericin B: 0.1~0.2g

Gentamicin: 0.07~0.13g

Reduced glutathione: 3-5mmol

Adenosine: 4-8mmol

Deionized water: 1000ml

Containing there are three types of antibiotic in ingredient:

Penicillin G sodium salt: being one kind of penicillin, refers in molecule containing penam, can destroy the cell wall one of bacterium Class antibiotic is had an effect in the bacterial reproduction phase, is the antibiotic by extracting in Penicillium notatum.

Amphotericin B: being polyene macrolide antibiotics, be produced by actinomyces and fungi and containing conjugated double bond and Antifungal antibiotic.The feature of polyenic compounds is all presented, polyene antibiotic mainly inhibits the cholesterol of fungal cell membrane, example Such as the cell membrane of fungi and mycoplasma.

Gentamicin: being one kind of aminoglycoside antibiotics, is to be connected by amino sugar and aminocyclitol by oxygen bridge At, mainly inhibition bacterium synthetic proteins, inhibit synthesis initial composite object, Induction of bacterial resultant fault albumen, at the same interfere The release of synthetic proteins, so as to cause bacterial death.

The preparation method of liquid is saved according to the adipose tissue of appeal, it is characterised in that:

Step 1, according to preparation step, weigh each constituent；

Step 2 is dissolved in deionized water, obtains saving liquid；

Step 3, by the packing of every part of preservation liquid, vacuum seal and cryo-conservation.

Further technical solution is that first each ingredient is dissolved in the deionized water of 100ml；Then 0.22 μm of filter mistake is utilized Filter obtains filtered fluid；Filtered fluid is finally diluted to 1000ml to obtain saving liquid.

The above operation is required to aseptically operate.

Benefit of the invention is as follows:

It joined penicillin G sodium salt, gentamicin, amphotericin B in preservation liquid of the present invention, fatty group can be prevented It is woven in during preservation by bacterium dirt

Specific embodiment

Combined with specific embodiments below the present invention is made further to explain.

Adipose tissue used in the following embodiment is the fat that patient abdomen is derived from operation.

Examples 1 to 3 is the preparation method that adipose tissue saves liquid, and each ingredient has a certain difference, is specifically shown in Table 1.

Table 1

Ingredient	Embodiment 1	Embodiment 2	Embodiment 3
				D-40-40	50g	60g	70g
NaCl	50mmol	55mmol	60mmol
				KCl	20mmol	25mmol	30mmol
MgSO4	4mmol	6mmol	8mmol
				Sucrose	40mmol	55mmol	60mmol
Penicillin G sodium salt	0.1g	0.15g	0.2g
				Amphotericin B	0.1g	0.15g	0.2g
Gentamicin	0.07g	0.1g	0.13g
				Reduced glutathione	3mmol	4mol	5mmol
Adenosine	4mol	6mol	8mmol
				Deionized water	1000ml	1000ml	1000ml

Adipose tissue in Examples 1 to 3 saves liquid, comprising the following steps:

Step 1 weighs each component by regulation；

Each ingredient is first dissolved in the deionized water of 100ml by step 2；Then filtering is obtained by filtration using 0.22 μm of filter Liquid；Filtered fluid is finally diluted to 1000ml to obtain saving liquid.

Step 3 will save liquid packing, be dispensed with the liquid storage bottle of 125ml, each liquid storage bottle is equipped with the preservation liquid of 100ml.

Step 5, vacuum and low temperature save, and the preservation liquid after packing is tightened bottle cap, and sealed membrane seals, after being packed into valve bag, It is reloaded into vacuum seal pocket, carries out vacuum seal.The preservation liquid of sealing is finally placed in -20 DEG C of preservations.

It is by the method that adipose tissue is stored in preservation liquid:

Step A, the preservation liquid of -20 DEG C of the defrosting at 4 DEG C.

Step B, the adipose tissue that will be obtained after operation is placed in 4 DEG C of preservation liquid at once and saves.

Step C, adipose tissue is saved into 7d.

Step D, step C treated adipose tissue is taken out, culture, observes the culture situation of fat cell.

According to Examples 1 to 3 method, every group of 100 parts of preservation liquid.The adipose tissue that will be obtained after operation, it is in vitro after at once It is placed in 4 DEG C of fat preservation liquid and tentatively saves.Adipose tissue is saved into 7d.It is finally to take out, cultivate to adipose tissue, observation The culture situation of fat tissue cell, living cell counting ratio.

It the results are shown in Table 2:

Table 2

	Embodiment 1	Embodiment 2	Embodiment 3
				Save sum	100	100	100
Survive sum	96	91	90
				Survival rate	96%	91%	90%

As shown in table 2, after adipose tissue saves in saving liquid, survival rate is high.Wherein the condition in embodiment 1 is most Excellent configuration method.

Above description is explanation of the invention, is not intended to limit the invention, and limited range of the present invention is referring to right It is required that the present invention can make any type of modification without prejudice to basic structure of the invention.

Claims

1. the invention discloses a kind of human fat tissues to save liquid, the fat of every 1000ml saves liquid and includes: