CN1188674A - Method for preparing tumour vaccinum - Google Patents
Method for preparing tumour vaccinum Download PDFInfo
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- CN1188674A CN1188674A CN97105061A CN97105061A CN1188674A CN 1188674 A CN1188674 A CN 1188674A CN 97105061 A CN97105061 A CN 97105061A CN 97105061 A CN97105061 A CN 97105061A CN 1188674 A CN1188674 A CN 1188674A
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Abstract
A process for preparing tumor vaccine includes such steps as taking paraffin wrapped cancer tissue section, preparing single-cell suspension, washing with buffer liquid, addition of trypsin, treating in microwave oven or with garlicin or aloe extract liquid, washing with buffer liquid, addition of immune adjuvant, and adding aldehyde fixating agents, and features easily available antigen, simple process, low cost, thorough deactivation, high safety and distinct curative effect.
Description
The invention belongs to the antitumor immune technical field, relate in particular to a kind of method of utilizing the paraffin embedding cancerous tissue to prepare tumor vaccine.
Use the tumour-specific active immunity treatment of tumor vaccine active immunity, for new approach has been opened up in the treatment of malignant tumor with excitating organism internal specific antineoplastic immune power.Because the complexity of tumor antigen, the research of allosome tumor vaccine does not make great progress as yet, and the autologous tumor vaccine then becomes one of research focus in current this area.In the prior art, the preparation of autologous tumor vaccine all is to adopt the cancerous cell of fresh cancerous tissue and work as antigenic source.For example Chinese patent application discloses the method that the fresh cancerous tissue of a kind of usefulness prepares tumor vaccine 93112060, No. 8.But,, can't in treatment, be used widely as the tumor vaccine in antigen source with fresh cancerous tissue, because be difficult to obtain the fresh tumor tissues of patient this moment from body to postoperative recurrence, transfer patient owing to be subjected to the restriction of preservation condition and holding time.If want the above-mentioned tumor vaccine of long preservation, it must be placed cryogenic conditions for a long time, this can cause the aldehydes polymerization, the antigen aldehyde radicalization, thus cover antigen, reduce tumor vaccine effectiveness greatly.In addition, adopt fresh tumor tissues to prepare tumor vaccine,, be difficult to sometimes the thorough deactivation of cancerous cell owing to be subjected to the condition effect such as time, temperature of the effectiveness of Drug resistance, deactivation medicine of cancerous cell and deactivation operation.
Purpose of the present invention will overcome the deficiency of above-mentioned prior art exactly, the tumor vaccine preparation method of provide a kind of antigen to originate easily, preparation method is easy, with low cost, be widely used, the tumor vaccine deactivation is thorough, safe in utilization, evident in efficacy.
The object of the present invention is achieved like this: a kind of method for preparing tumour vaccinum is characterized in that taking the following step:
(1) gets the section of paraffin embedding cancerous tissue,, make single cell suspension through dewaxing, aquation, digestion process;
(2) wash with buffer;
(3) add trypsin and put 3 minutes, or put into microwave oven and handled 10 minutes;
(4) wash with buffer;
(5) add immunological adjuvant;
(6) add aldehyde fixative.
Purpose of the present invention can also realize like this: a kind of method for preparing tumour vaccinum is characterized in that taking the following step:
(1) gets the section of paraffin bag reason cancerous tissue,, make single cell suspension through dewaxing, aquation, digestion process;
(2) wash with buffer;
(3) hatched 20～30 minutes with garlicin or Aloe extraction solution;
(4) wash with buffer;
(5) add immunological adjuvant;
(6) add aldehyde fixative.
In order to realize purpose of the present invention better, the used adjuvant of above-mentioned each method is bacillus calmette-guerin vaccine or PPD.
Tumor vaccine preparation method of the present invention, the antigen source is easy, and the patient generally is long-term conventional the file from paraffin body embedding cancerous tissue at its operation hospital pathology department, and the cancerous tissue of depositing is enough to the expense of satisfied preparation tumor vaccine; Preparation method is easy, grasps easily; With low cost, do not need special equipment, be convenient to the less hospital application of basic unit's scale; Can be used for all postoperative tumor patients,, be not subjected to time restriction with using with system; Because the antigen source is the paraffin embedding cancerous tissue, so deactivation is definitely thorough, safe and reliable; The tumor vaccine of preparation has significantly anticancer and metastasis effect, energy excitating organism specificity antineoplastic cellular immunization and humoral immunization, and curative effect is remarkable especially.
Describe embodiments of the invention below in detail.
Example 1.
Get the thick paraffin embedding cancerous tissue section of 50 μ m, immersed in the dimethylbenzene 12 hours, change in the dehydrated alcohol 12 hours then over to, immerse 95% ethanol more successively, 75% ethanol, in the PBS liquid each 30 minutes (in 75% ethanol, taking out the back) by strict sterile working, placing temperature then is 37 ℃ 0.14% collagenase 30 minutes, reuse 100 order nickel screens filter, make single cell suspension, then with PBS liquid washing 3 times, the trypsin of adding 1% was put 3 minutes or was put into microwave oven and handled 10 minutes, and reuse PBS liquid washing 3 times adds bacillus calmette-guerin vaccine or PPD, add dialdehyde or 0.03% formaldehyde, the packing of peace bottle at last of 0.025% penta again.
Example 2.
Get the thick paraffin embedding cancerous tissue section of 50 μ m, immersed in the dimethylbenzene 12 hours, change in the dehydrated alcohol 12 hours then over to, immerse 95% ethanol more successively, 75% ethanol, in the PBS liquid each 30 minutes (in 75% ethanol, taking out the back) by strict sterile working, placing temperature then is 37 ℃ 0.14% collagenase 30 minutes, reuse 100 order nickel screens filter, make single cell suspension, then with PBS liquid washing 3 times, the trypsin of adding 1% was put 3 minutes or was put into microwave oven and handled 10 minutes, reuse PBS liquid washing 3 times, hatched under 37 ℃ 20～30 minutes with 0.5～3% garlicin or Aloe extraction solution then, reuse PBS liquid washing 3 times adds bacillus calmette-guerin vaccine or PPD, add 0.025% glutaraldehyde or 0.03% formaldehyde again, at last the packing of peace bottle.
(the tumor strain is that 615 mouse mastopathy cell plant height lungs shift inferior strain: Ca761-FP8/L) to the experimental data of lotus tumor 615 mices treatment to provide tumor vaccine that the foregoing description makes below.Processing method number of animals tumor weight (X ± SD gram) lung shifts number time-to-live (X ± SD days) example 18 0.2945 ± 0.299 3 50.505 ± 11.780 example 28 0.2517 ± 0.340 2 51.608 ± 10.975 nothing 8 0.5769 ± 0.275 7 36.500 ± 6.950
Claims (3)
1. method for preparing tumour vaccinum is characterized in that taking the following step:
(1) gets the section of paraffin embedding cancerous tissue,, make single cell suspension through dewaxing, aquation, digestion process;
(2) wash with buffer;
(3) add trypsin and put 3 minutes, or put into microwave oven and handled 10 minutes;
(4) wash with buffer;
(5) add immunological adjuvant;
(6) add aldehyde fixative.
2. method for preparing tumour vaccinum is characterized in that taking the following step:
(1) gets the section of paraffin bag reason cancerous tissue,, make single cell suspension through dewaxing, aquation, digestion process;
(2) wash with buffer;
(3) hatched 20～30 minutes with garlicin or Aloe extraction solution;
(4) with the buffer washing;
(5) add immunological adjuvant;
(6) add aldehyde fixative.
3. method for preparing tumour vaccinum according to claim 1 and 2 is characterized in that used adjuvant is bacillus calmette-guerin vaccine or PPD.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97105061A CN1100569C (en) | 1997-01-24 | 1997-01-24 | Method for preparing tumour vaccinum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97105061A CN1100569C (en) | 1997-01-24 | 1997-01-24 | Method for preparing tumour vaccinum |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1188674A true CN1188674A (en) | 1998-07-29 |
| CN1100569C CN1100569C (en) | 2003-02-05 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN97105061A Expired - Fee Related CN1100569C (en) | 1997-01-24 | 1997-01-24 | Method for preparing tumour vaccinum |
Country Status (1)
| Country | Link |
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| CN (1) | CN1100569C (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN149012B (en) * | 1978-04-20 | 1981-08-15 | Duncan Lee Mccollester |
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1997
- 1997-01-24 CN CN97105061A patent/CN1100569C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1100569C (en) | 2003-02-05 |
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