CN101892219A - Application of static constant magnetic field for prolonging hold time of primary cell - Google Patents
Application of static constant magnetic field for prolonging hold time of primary cell Download PDFInfo
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- CN101892219A CN101892219A CN 201010226998 CN201010226998A CN101892219A CN 101892219 A CN101892219 A CN 101892219A CN 201010226998 CN201010226998 CN 201010226998 CN 201010226998 A CN201010226998 A CN 201010226998A CN 101892219 A CN101892219 A CN 101892219A
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Abstract
The invention discloses application of static constant magnetic field used for prolonging hold time of primary cell, including that cells are placed in a magnetic field to be cultured and the magnetic field is arranged on the outer wall close to the cells. Concretely, the magnetic field is arranged at the bottom of a cell culture plate when the cell is cultured by adopting the cell culture plate; the magnetic filed is arranged at the bottom of a cell culture bottle when the cell is cultured by adopting the cell culture bottle; and the magnetic field is arranged on the outer wall of a roller bottle when the cell is cultured by adopting the roller bottle. The magnetic field is a static constant one, and magnetic force is 500-3000 Gauss. The primary cell can be histocytes coming from all animals including human, such as hepatocyte, nephrocyte, lens cell and testicular cell. The invention adopts a manner of adding a magnetic field into the cell culture environment to prolong hold time of primary cell in vitro, the hold time of cell can be more than 30 days, and cell growth is vigorous, thus effect is good.
Description
Technical field
The present invention relates to the application of static constant magnetic field in the prolongation primary cell is held time, belong to field of cell culture.
Background technology
It is multi-field research tools commonly used such as biology, medical science that cells in vitro is cultivated, wherein diploid cell, especially the cultivation of primary cell occupies critical positions in association area.At present, most primary cells go down to posterity and cultivate difficulty, even can not go down to posterity (human embryo lung (HEL) is external to go down to posterity for 50～60 generations), and it is 10～20d that the primary cell of general vitro culture is held time, and then cell aging for a long time loses using value again.Therefore, people are badly in need of a kind ofly can prolonging the method that cell is held time.
In the prior art, in order to prolong the primary cell survival time, person skilled takes to reduce culture temperature more, reduce nutritive ingredient in the nutrient solution, and (nutrient solution that reduces nutritive ingredient is called and keeps liquid, such as the MEM substratum that contains 2% new-born calf serum) etc. method, do not see that useful static constant magnetic field prolongs the report that primary cell is held time.
Summary of the invention
At above-mentioned prior art, be the primary cell that the solves vitro culture short defective of holding time, the invention provides a solution, that is: utilize magnetic field to prolong holding time of primary cell, method of the present invention can make cell hold time above 30d, and the cell growing way is vigorous, even along the wall bulk-growth.
The mode of using is: place magnetic field to cultivate in cell, magnetic field is arranged on the outer wall of cell.
Be specially:, then magnetic field is arranged at the bottom of Tissue Culture Plate for adopting the Tissue Culture Plate cultured cells; For the cell that adopts Tissue Culture Flask, then magnetic field is arranged at the bottom of Tissue Culture Flask; For adopting the rolling bottle cultured cells, then magnetic field is arranged on the rolling bottle outer wall and (does not hinder rotation).
Described magnetic field is static constant magnetic field, and the magnetic force scope is 500～3000 Gausses.
Described primary cell is to derive from the histocyte that comprises all human animals, such as liver cell, nephrocyte, crystal cell, testicular cell etc.
Attention: method of the present invention is not suitable for tumour cell, because tumour cell often shows as the change of growth pattern.
The mode extension body exogenesis in the present invention's employing interpolation magnetic field in cell culture environment is held time for cell, cell is held time above 30d, and the cell growing way is vigorous, and effect is very good.
Description of drawings
Fig. 1 is human embryonic lung cell's cultivation figure (200 *).
Embodiment
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1 human embryonic lung cell's cultivation:
1: former generation human embryonic lung cell's preparation: get the embryo lung tissue under the aseptic condition, (local laminar flow) shreds with scissors in the super clean bench, add pancreatin and under 37 ℃ of conditions, digest 20min, abandon pancreatin, add cell nutrient solution suspension piping and druming, then suspension is crossed 100 order gauzes and removed tissue block, it is 10 that filtrate is adjusted cell concn
5/ ml is inoculated in the Tissue Culture Flask, puts static cultivation in 37 ℃, 5% carbonic acid gas, 95% air ambient, grows up to individual layer after 3 days, is former generation human embryonic lung cell.
2: the human embryonic lung cell's goes down to posterity
The human embryonic lung cell who grows up to individual layer is washed 3 times with balanced salt, remove not attached cell and tissue block, add 0.125% pancreatin and digest 3min at ambient temperature, abandon pancreatin, add cell nutrient solution piping and druming and suspend, divide bottle to go down to posterity with 1: 2 ratio then, put static cultivation in 37 ℃, 5% carbonic acid gas, 95% air ambient, grow up to individual layer after 3 days, for inferior for the human embryonic lung cell.
3: the human embryonic lung cell keeps
Abandon nutrient solution with growing up to the inferior of individual layer for human embryonic lung cell's (4 bottles), adding the MEM that contains 2% new-born calf serum is kept, insert 37 ℃ for 2 bottles, 5% carbonic acid gas, static cultivation in the environment of the static magnetic field of 95% air (800 Gauss), simultaneously put no magnetic environment and organize (surplus condition is identical) in contrast for 2 bottles, observation of cell situation during 20d (middle do not change or add new nutrient solution), find that all cells nutrient solution has become lemon yellow, show that cell is in sour environment, 2 bottles of cells of control group become sparse, swell, the opacifying property particle occurs, and 2 bottles of cellular fories are good in the magnetic field, continue to keep termination test when being cultured to 35d, observe nutrient solution and be yellow (it is acid that pH is), the microscopically observation of cell is fibrous, and tiling is arranged and is fine and close individual layer, does not see obvious opacifying property particle, nothing comes off, and sees Fig. 1.Show that the static magnetic field environment has prolonged the cell survival time.
Claims (6)
1. the application of static constant magnetic field in the prolongation primary cell is held time.
2. application according to claim 1 is characterized in that, application mode is: place magnetic field to cultivate in cell, magnetic field is arranged on the outer wall of cell.
3. application according to claim 2 is characterized in that: for adopting the Tissue Culture Plate cultured cells, then magnetic field is arranged at the bottom of Tissue Culture Plate; For the cell that adopts Tissue Culture Flask, then magnetic field is arranged at the bottom of Tissue Culture Flask; For adopting the rolling bottle cultured cells, then magnetic field is arranged on the rolling bottle outer wall.
4. according to claim 1 or 2 or 3 described application, it is characterized in that: described magnetic field is permanent-magnetic field, and magnetism intensity is 500～3000 Gausses.
5. according to claim 1 or 2 or 3 described application, it is characterized in that: described primary cell is adherent diploid cell.
6. application according to claim 5 is characterized in that: described primary cell is the human embryonic lung cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010226998 CN101892219A (en) | 2010-07-15 | 2010-07-15 | Application of static constant magnetic field for prolonging hold time of primary cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010226998 CN101892219A (en) | 2010-07-15 | 2010-07-15 | Application of static constant magnetic field for prolonging hold time of primary cell |
Publications (1)
| Publication Number | Publication Date |
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| CN101892219A true CN101892219A (en) | 2010-11-24 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN 201010226998 Pending CN101892219A (en) | 2010-07-15 | 2010-07-15 | Application of static constant magnetic field for prolonging hold time of primary cell |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788080A (en) * | 2003-05-14 | 2006-06-14 | 株式会社日本组织工程 | Cell culture method and cultured tissue |
| CN101463319A (en) * | 2009-01-07 | 2009-06-24 | 中国医学科学院生物医学工程研究所 | Apparatus for cell cultivation by pulse electromagnetic field |
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2010
- 2010-07-15 CN CN 201010226998 patent/CN101892219A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788080A (en) * | 2003-05-14 | 2006-06-14 | 株式会社日本组织工程 | Cell culture method and cultured tissue |
| CN101463319A (en) * | 2009-01-07 | 2009-06-24 | 中国医学科学院生物医学工程研究所 | Apparatus for cell cultivation by pulse electromagnetic field |
Non-Patent Citations (1)
| Title |
|---|
| 《中华理疗杂志》 20001231 王卫国等 头皮下植入磁铁并脉冲电治疗耳鸣观察 第186-187页 1-6 第23卷, 第3期 2 * |
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Open date: 20101124 |