CN109762780A - A kind of separation of people's alveolar epithelial cells and cultural method - Google Patents
A kind of separation of people's alveolar epithelial cells and cultural method Download PDFInfo
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Abstract
The present invention provides a kind of separation of people's alveolar epithelial cells and cultural methods, and the group of acquisition, which is woven under the encirclement of activated base, can be effectively performed adherent growth, and serum composition can inhibit the division and differentiation of alveolar epithelial cells, to substantially increase cell viability；Fragment of tissue shake culture under specific gaseous environment, it is possible to prevente effectively from the tissue samples of acquisition cause to damage due to ischemic, in favor of subsequent culture operation；Allantoin, creatine and the asiaticosid added in secondary culture base can effectively improve cell viability and splitting ability, so that cell be promoted to grow and be proliferated；Using 3T3 cell as trophoderm, good growth substrate can be provided for primary alveolar epithelial cells, the growth division of alveolar epithelial cells be effectively facilitated, to greatly improve culture effect.By the above method, the activity and proliferation rates of alveolar epithelial cells can be significantly improved, to provide good experimental material for correlative study project.
Description
Technical field
The invention belongs to tissue engineering technique field, in particular to a kind of people's alveolar epithelial cells separation and cultural method.
Background technique
Human airway epithelial cells are the active cells of function, do not merely comprise the physical barriers of respiratory tract, are also had to chemically
The conversion metabolism of poisonous substance or drug, has the function of autocrine or paracrine, simultaneously participates in the regulation to adjacent cells function.
Just in vitro primary cell remains with original biological heredity characteristic, can the growth of antimer inner cell general features, therefore
It is studied suitable for drug and therapy etc..Establish the cell in vitro mould of the alveolar epithelial cells of normal alveolar epithelial cells and lesion
Type is conducive to the pathological change for being best understood from alveolar epithelial cells, also more easy, reliable compared with for zoopery.But mesh
It is preceding still immature to the in vitro culture of alveolar epithelial cells.Therefore, explore it is a kind of can improve alveolar epithelial cells activity and proliferation
The extracorporeal culturing method of ability, it has also become those skilled in the art's technical problem urgently to be solved.
Summary of the invention
In order to solve the above-mentioned technical problems, the present invention provides a kind of separation of people's alveolar epithelial cells and cultural methods.
Specific technical solution of the present invention is as follows:
The present invention provides a kind of separation of people's alveolar epithelial cells and cultural methods, include the following steps:
S 1: lobe of the lung sample is aseptically acquired, tracheae is cut into lamellar structure, obtains the group of alveolar epithelium tissue
Knit fragment；
S2: being configured to the activated base of cell adherent growth, is coated on culture dish and is solidified；
S3: the setting inoculation position on the cured activated base, and the fragment of tissue is inoculated into the inoculation
On position, the tissue pieces is made to be attached to culture dish bottom wall；
S4: adding activation culture liquid into the fragment of tissue after inoculation, is placed in mixture of oxygen and carries out activation culture；
The step is it is possible to prevente effectively from the tissue samples of acquisition cause to damage due to ischemic, in favor of subsequent culture behaviour
Make；
S5: the fragment of tissue Jing Guo activation culture being transferred in the new culture dish for being coated with activated base, and addition is former
It is commissioned to train nutrient solution, originally culture is carried out in CO2 incubator, cultivate to be transferred to fragment of tissue after 8~10d and new be coated with activation
In the culture dish of substrate, repeats originally culture process several times, obtain primary cell；
S6: by the primary cell with every bottle 104The concentration of a cell is inoculated with, using the 3T3 cell by irradiation
Make trophoderm, secondary culture liquid is added and carries out suspension culture, obtains passage cell；
Using 3T3 cell as trophoderm, good growth substrate can be provided for primary alveolar epithelial cells；By primary lung
After steeping epithelial cell inoculation thereon, alveolar epithelial cells can obviously inhibit the growth of 3T3 cell, and own growth is good,
And 3T3 cell is constantly pushed into culture dish edge.By the above method, the growth point of alveolar epithelial cells can be effectively facilitated
It splits, to greatly improve culture effect.
Further, the specific method is as follows by step S1:
Lobe of the lung sample is aseptically acquired, is immersed in L-15 culture solution, tracheostomize, and be cut into 1cm2Square
Sheet obtains the fragment of tissue of alveolar epithelium tissue.
Further, the specific method is as follows by step S2:
It is configured to the activated base of cell adherent growth, takes 1ml to be coated on 6cm culture dish bottom surface, is placed in 36.5 DEG C
2h is placed in CO2 incubator, solidifies the activated base, and 5ml activation culture liquid is added.
Further, the specific method is as follows by step S3:
1cm is depicted with scalpel at culture dish bottom surface edge2Square, and remove activated base therein,
Form inoculation position；The fragment of tissue epithelial surface is moved up into the inoculation position, activation culture liquid is removed, in room temperature
3~5min of lower standing, makes tissue block be attached to culture dish bottom wall.
Further, the specific method is as follows by step S4:
Into the fragment of tissue after inoculation add 3ml DMEM culture solution, be put into can air control chamber, into gas chamber inject mixing
Gas chamber is placed on shaking table by oxygen, is cultivated for 24 hours under the conditions of 36.5 DEG C, 10r/min, flows culture solution from fragment of tissue surface
It crosses, sufficiently infiltrate fragment of tissue；Then DMEM culture solution and mixture of oxygen are replaced, 6~8d of culture, every primary training of 2d replacement are continued
Nutrient solution and mixture of oxygen；
The mixture of oxygen includes 50%O2, 45%N2And 5%CO2。
Above-mentioned mixture of oxygen is it is possible to prevente effectively from the tissue samples of acquisition cause to damage due to ischemic, in favor of subsequent
Culture operation.
Further, the specific method is as follows by step S5:
S5.1: vacuum attraction is carried out with new 10cm culture dish protrusion activated base, and to activated base bottom, with dissection
Graduating with cutter draws new inoculation position, and fragment of tissue is transferred in new culture medium, adds aseptic culture fluid, places 3 at room temperature
~5h；
S5.2: being added 10ml LHC-9 culture solution (be purchased from ThermoFisher), in the 5%CO2 incubator of humidifying into
Row originally culture, every 3~4d replace culture solution；
S5.3: after 8~10d of culture, tissue block is moved into the new culture dish for being coated with activated base, repeats step
S5.2 several times, obtains primary cell.
Further, the specific method is as follows by step S6:
S6.1: washing the primary cell with PBS buffer solution, and digested with digestive juice, after digestion,
With culture solution suspension cell and count；According to every bottle 104A inoculum concentration is inoculated into new culture bottle；
S6.2: being added the 3T3 cell irradiated by Co60 of 5~10 times of quantity, secondary culture liquid be added after mixing,
It is placed at 37 DEG C and cultivates 15~30 days to get passage cell is arrived；
The processing method of 3T3 cell is as follows:
(1) with the DMEM culture medium culture 3T3 cell containing 10% calf serum, culture solution is replaced after 48h, continues to cultivate
15~30 days；
(2) it is irradiated by 60Gy Co60,3T3 cell is inactivated；
(3) it is digested with 3T3 cell of the digestive juice to inactivation, cleans two with the DMEM culture medium without serum
It is secondary, and be suspended.
Co60 irradiation can make 3T3 cell loss splitting ability, while not influence the activity of original cell, to be primary
The trophoderm of alveolar cell offer excellent.
Further, the activated base is the LHC-9 culture solution added with following ingredient:
The viscous element 0.2% of people's fibre；Collagen 1%；Fetal calf serum 12%.
People's fibre, which glues element, can promote cell adherent growth；Collagen can effective coated cell, protection cell；Serum can press down
The division and differentiation of alveolar epithelial cells processed, in the form of keeping cell and vigor.Mentioned component can be improved cell viability, promote
Into cell adherent growth.
Further, the digestive juice is 1% polyvinyl pyrrole added with 0.02% trypsase and 0.02%EDTA
Alkanone solution.
Mentioned component can effectively digest adherent cell, to keep cell fully dispersed, in favor of subsequent
Secondary culture.
Further, the secondary culture liquid is the DMEM culture medium added with following ingredient:
Sodium lactate Green 8~12%, allantoin 1.5~2.5%, creatine 0.5~1%, asiaticosid 0.2~0.5% with
And chlorogenic acid 0.1~0.2%.
Sodium lactate Green is the sterile water solution of sodium lactate, sodium chloride, potassium chloride and calcium chloride, can be used for providing close to people
The moderate environment of the osmotic pressure of body, in favor of the survival of cell；Allantoin can promote cell growth, cutin-softening albumen, from
And accelerate tissue update and wound healing；Creatine can quickly provide energy for cell metabolism, promote cell growth and update；Product
Asiaticoside can accelerate cell metabolism and division, promote tissue growth and wound healing；Chlorogenic acid has excellent antibacterial, disease-resistant
Toxic effect fruit, and can effectively anti-oxidant, promotion cell activity.Using the secondary culture liquid of said ratio, can effectively improve thin
Born of the same parents' vigor promotes cell division, to be quickly obtained good passage cell.
Beneficial effects of the present invention are as follows: the present invention provides a kind of separation of people's alveolar epithelial cells and cultural methods, adopt
The group of collection, which is woven under the encirclement of activated base, can be effectively performed adherent growth, and serum composition can inhibit alveolar epithelial cells
Division and differentiation, to substantially increase cell viability；Fragment of tissue shake culture under specific gaseous environment, can be effective
The tissue samples of acquisition are avoided to cause to damage due to ischemic, in favor of subsequent culture operation；It is added in secondary culture base
Allantoin, creatine and asiaticosid can effectively improve cell viability and splitting ability, so that cell be promoted to grow and be proliferated；It will
3T3 cell can provide good growth substrate for primary alveolar epithelial cells, it is thin to effectively facilitate alveolar epithelium as trophoderm
The growth of born of the same parents is divided, to greatly improve culture effect.By the above method, the activity of alveolar epithelial cells can be significantly improved
And proliferation rates, to provide good experimental material for correlative study project.
Detailed description of the invention
Fig. 1 is the electromicroscopic photograph of alveolar epithelial cells；
Fig. 2 is the growth curve cultivated using distinct methods alveolar epithelial cells.
Specific embodiment
The preparation of main agents and culture medium
A.L-15 culture medium: specific ingredient is as follows:
10% fetal calf serum, 900mg/L D- galactolipin, 300mg/L L-Glutamine, 550mg/L Sodium Pyruvate.
B.DMEM culture medium: specific ingredient is as follows:
C.PBS buffer:
Weigh 8g NaCl, 0.2g KCl, 1.44g Na2HPO4With 0.24g KH2PO4, it is dissolved in 800mL distilled water, uses
HCl adjusts the pH value of solution to 7.4, and finally plus distilled water is settled to 1L.In 15lbf/in2 (1034 × 105Pa) high pressure
Lower steam sterilization (at least 20min) is stored in room temperature or 4 DEG C of refrigerators.
Below with reference to following embodiment, invention is further described in detail.
Embodiment 1
A kind of separation of people's alveolar epithelial cells and cultural method, include the following steps:
S1: lobe of the lung sample is aseptically acquired, tracheae is cut into lamellar structure, obtains the tissue of alveolar epithelium tissue
Fragment；
S2: being configured to the activated base of cell adherent growth, is coated on culture dish and is solidified；
S3: the setting inoculation position on the cured activated base, and the fragment of tissue is inoculated into the inoculation
On position, the tissue pieces is made to be attached to culture dish bottom wall；
S4: adding activation culture liquid into the fragment of tissue after inoculation, is placed in mixture of oxygen and carries out activation culture；
S5: the fragment of tissue Jing Guo activation culture being transferred in the new culture dish for being coated with activated base, and addition is former
It is commissioned to train nutrient solution, in CO2It carries out originally culture in incubator, cultivates to be transferred to fragment of tissue after 8~10d and new be coated with activation
In the culture dish of substrate, repeats originally culture process several times, obtain primary cell；
S6: by the primary cell with every bottle 104The concentration of a cell is inoculated with, using the 3T3 cell by irradiation
Make trophoderm, secondary culture liquid is added and carries out suspension culture, obtains passage cell.
Embodiment 2
A kind of separation of people's alveolar epithelial cells and cultural method, including each step that embodiment 1 provides, wherein step S1
The specific method is as follows:
Lobe of the lung sample is aseptically acquired, is immersed in L-15 culture solution, tracheostomize, and be cut into the square of 1cm2
Sheet obtains the fragment of tissue of alveolar epithelium tissue.
The specific method is as follows by step S2:
It is configured to the activated base of cell adherent growth, takes 1ml to be coated on 6cm culture dish bottom surface, is placed in 36.5 DEG C
CO22h is placed in incubator, solidifies the activated base, and 5ml activation culture liquid is added.
The specific method is as follows by step S3:
1cm is depicted with scalpel at culture dish bottom surface edge2Square, and remove activated base therein,
Form inoculation position；The fragment of tissue epithelial surface is moved up into the inoculation position, activation culture liquid is removed, in room temperature
3~5min of lower standing, makes tissue block be attached to culture dish bottom wall.
The specific method is as follows by step S4:
Into the fragment of tissue after inoculation add 3ml DMEM culture solution, be put into can air control chamber, into gas chamber inject mixing
Oxygen (50%O2, 45%N2And 5%CO2), gas chamber is placed on shaking table, is cultivated for 24 hours under the conditions of 36.5 DEG C, 10r/min,
Then DMEM culture solution and mixture of oxygen are replaced, 6~8d of culture is continued, every 2d replaces a culture solution and mixture of oxygen.
The specific method is as follows by step S5:
S5.1: vacuum attraction is carried out with new 10cm culture dish protrusion activated base, and to activated base bottom, with dissection
Graduating with cutter draws new inoculation position, and fragment of tissue is transferred in new culture medium, adds aseptic culture fluid, places 3 at room temperature
~5h；
S5.2: 10ml LHC-9 culture solution is added, in the 5%CO of humidifying2Originally culture, every 3~4d are carried out in incubator
Replace culture solution；
S5.3: after 8~10d of culture, tissue block is moved into the new culture dish for being coated with activated base, repeats step
S5.2 several times, obtains primary cell.
The specific method is as follows by step S6:
S6.1: primary cell is washed with PBS buffer solution, and is digested with digestive juice；After digestion, with training
Nutrient solution suspension cell simultaneously counts；According to every bottle 104A inoculum concentration is inoculated into new culture bottle；
S62: the 3T3 cell irradiated by Co60 of 5~10 times of quantity is added, culture solution is added after mixing, is placed in
15~30 days are cultivated at 37 DEG C to get passage cell is arrived.
Wherein, the processing method of 3T3 cell is as follows:
(1) with the DMEM culture medium culture 3T3 cell containing 10% calf serum, culture solution is replaced after 48h, continues to cultivate
15~30 days；
(2) it is irradiated by 60Gy Co60,3T3 cell is inactivated；
(3) it is digested with 3T3 cell of the digestive juice to inactivation, is cleaned twice with the DMEM culture medium without serum, and
It is suspended.
Embodiment 3
A kind of separation of people's alveolar epithelial cells and cultural method, including each step that embodiment 2 provides, wherein the work used
Change substrate is the LHC-9 culture solution added with following ingredient:
The viscous element 0.2% of people's fibre；Collagen 1%；Fetal calf serum 12%.
Embodiment 4
A kind of separation of people's alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein what is used is described
Digestive juice is 1% polyvinylpyrrolidonesolution solution added with 0.02% trypsase and 0.02%EDTA.
Embodiment 5
A kind of separation of alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein the passage training used
Nutrient solution is the DMEM culture medium added with following ingredient:
Sodium lactate Green 8%, allantoin 1.5%, creatine 1%, asiaticosid 0.2% and chlorogenic acid 0.2%.
Embodiment 6
A kind of separation of alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein the activation base used
Bottom is the LHC-9 culture solution added with following ingredient:
The viscous element 0.2% of people's fibre；Collagen 1%；Fetal calf serum 12%；
The digestive juice used is 1% polyvinylpyrrolidone added with 0.02% trypsase and 0.02%EDTA
Solution；
The secondary culture liquid used is the DMEM culture medium added with following ingredient:
Sodium lactate Green 12%, allantoin 2.5%, creatine 0.5%, asiaticosid 0.5% and chlorogenic acid 0.1%.
Reference examples 1
A kind of separation of alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein the passage training used
Nutrient solution is the DMEM culture medium added with following ingredient:
Sodium lactate Green 8%, allantoin 2.5%, creatine 0.5% and chlorogenic acid 0.2%
Reference examples 2
A kind of separation of alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein the passage training used
Nutrient solution is the DMEM culture medium added with following ingredient:
Sodium lactate Green 12%, fetal calf serum 3%, asiaticoside 0.5% and chlorogenic acid 0.1%.
Reference examples 3
A kind of separation of alveolar epithelial cells and cultural method, including each step described in embodiment 2, wherein the passage training used
Nutrient solution is RPMI1640 culture medium.
Experimental example
Dyeing identification
The method that the offer of embodiment 2,4,5,6 is respectively adopted is experimental group 1~4, the method point provided with reference examples 1~3
Not as a control group 1~3, and with (the alveolar cell proliferation and Vimentin albumen that asiaticosid induces TGF-β 1 such as Liu Tao
Expression influences China Immunology impurity, 2019,35:25-29) cultural method as a control group 4 provided, respectively to it is same come
The alveolar epithelium tissue specimen in source is cultivated, and keratin antibody is dyed after culture, and is seen under Electronic Speculum
It examines, as a result as shown in Figure 1.
Electronic Speculum is the results show that the keratin positive rate of group of cells reaches 90% or more, it was demonstrated that group of cells is epithelium
Source property, i.e. alveolar epithelial cells.
Growth curve
Secondary culture is carried out to group of cells with 96 orifice plates, is respectively that group of cells draws growth curve using CCK-8 method,
Sample time is respectively the 1st of secondary culture the, 2,3,5,7,10,15d, as a result as shown in Figure 2.
By curve it is found that the cell quantity of experimental group each group and speedup are above control group each group, wherein with experimental group 4
Proliferation activity highest shows that method provided by the present application can significantly improve the reproduction activity of alveolar epithelial cells, especially with reality
It is best to test the method that 4 (i.e. embodiments 6) of group provide.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of people's alveolar epithelial cells separation and cultural method, which comprises the steps of:
S1: lobe of the lung sample is aseptically acquired, tracheae is cut into lamellar structure, the tissue for obtaining alveolar epithelium tissue is broken
Piece；
S2: being configured to the activated base of cell adherent growth, is coated on culture dish and is solidified；
S3: the setting inoculation position on the cured activated base, and the fragment of tissue is inoculated into the inoculation position
On, so that the tissue pieces is attached to culture dish bottom wall；
S4: adding activation culture liquid into the fragment of tissue after inoculation, is placed in mixture of oxygen and carries out activation culture；
S5: the fragment of tissue Jing Guo activation culture being transferred in the new culture dish for being coated with activated base, adds primary training
Nutrient solution, in CO2It carries out originally culture in incubator, cultivates to be transferred to fragment of tissue after 8~10d and new be coated with activated base
Culture dish in, repeat originally culture process several times, obtain primary cell；
S6: by the primary cell with every bottle 104The concentration of a cell is inoculated with, and is nourished using the 3T3 cell by irradiation
Layer is added secondary culture liquid and carries out suspension culture, obtains passage cell.
2. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S1
Method is as follows:
Lobe of the lung sample is aseptically acquired, is immersed in L-15 culture solution, tracheostomize, and be cut into 1cm2Square sheet,
Obtain the fragment of tissue of alveolar epithelium tissue.
3. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S2
Method is as follows:
It is configured to the activated base of cell adherent growth, takes 1ml to be coated on 6cm culture dish bottom surface, is placed in 36.5 DEG C of CO2Training
It supports and places 2h in case, solidify the activated base, and 5ml activation culture liquid is added.
4. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S3
Method is as follows:
The square of 1cm2 is depicted with scalpel at culture dish bottom surface edge, and removes activated base therein, is formed
It is inoculated with position；The fragment of tissue epithelial surface is moved up into the inoculation position, activation culture liquid is removed, it is quiet at room temperature
3~5min is set, tissue block is made to be attached to culture dish bottom wall.
5. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S4
Method is as follows:
Into the fragment of tissue after inoculation add 3ml DMEM culture solution, be put into can air control chamber, mixture of oxygen is injected into gas chamber,
Gas chamber is placed on shaking table, is cultivated for 24 hours under the conditions of 36.5 DEG C, 10r/min, HB culture solution and mixture of oxygen are then replaced, after
Continuous culture 6~8d, every 2d replace a culture solution and mixture of oxygen；
The mixture of oxygen is 50%O2, 45%N2And 5%CO2。
6. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S5
Method is as follows:
S5.1: carrying out vacuum attraction with new 10cm culture dish protrusion activated base, and to activated base bottom, with dissection graduating with cutter
New inoculation position is drawn, and fragment of tissue is transferred in new culture medium, sterile DMEM culture solution is added, places 3 at room temperature
~5h；
S5.2: 10ml LHC-9 culture solution is added, in the 5%CO of humidifying2Originally culture, every 3~4d replacement training are carried out in incubator
Nutrient solution；
S5.3: after 8~10d of culture, tissue block being moved into the new culture dish for being coated with activated base, if repeating step S5.2
Dry time, obtain primary cell.
7. people's alveolar epithelial cells separation as described in claim 1 and cultural method, which is characterized in that the specific side of step S6
Method is as follows:
S6.1: the primary cell is washed with PBS buffer solution, and is digested with digestive juice, after digestion, with training
Nutrient solution suspension cell simultaneously counts；According to every bottle 104A inoculum concentration is inoculated into new culture bottle；
S6.2: the 3T3 cell irradiated by Co60 of 5~10 times of quantity is added, secondary culture liquid is added after mixing, is placed in
15~30 days are cultivated at 37 DEG C to get passage cell is arrived；
The processing method of 3T3 cell is as follows:
(1) with the DMEM culture medium culture 3T3 cell containing 10% calf serum, replace culture solution after 48h, continue culture 15~
30 days；
(2) it is irradiated by 60Gy Co60,3T3 cell is inactivated；
(3) it is digested with 3T3 cell of the digestive juice to inactivation, is cleaned twice with the DMEM culture medium without serum, and
It is suspended.
8. such as people's alveolar epithelial cells according to any one of claims 1 to 7 separation and cultural method, which is characterized in that institute
Stating activated base is the LHC-9 culture solution added with following ingredient:
The viscous element 0.2% of people's fibre；Collagen 1%；Fetal calf serum 12%.
9. such as people's alveolar epithelial cells according to any one of claims 1 to 7 separation and cultural method, which is characterized in that institute
Stating digestive juice is 1% polyvinylpyrrolidonesolution solution added with 0.02% trypsase and 0.02%EDTA.
10. such as people's alveolar epithelial cells according to any one of claims 1 to 7 separation and cultural method, which is characterized in that institute
Stating secondary culture liquid is the DMEM culture medium added with following ingredient:
Sodium lactate Green 8~12%, allantoin 1.5~2.5%, creatine 0.5~1%, asiaticosid 0.2~0.5% and green
Ortho acid 0.1~0.2%.
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| CN108774629A (en) * | 2018-04-16 | 2018-11-09 | 中国科学院成都生物研究所 | A kind of primary culture method of Microhyla ornata alveolar epithelial cells |
| CN113913369A (en) * | 2021-11-25 | 2022-01-11 | 乐山市中医医院 | Selective medium for alveolar type I epithelial cells and separation and purification method thereof |
| CN113913369B (en) * | 2021-11-25 | 2022-07-01 | 四川大学华西医院 | Selective medium for alveolar type I epithelial cells and separation and purification method thereof |
| CN116814533A (en) * | 2023-08-28 | 2023-09-29 | 苏州冠佲生物医药科技有限公司 | Cell culture medium for in vitro maintenance of phenotype and function of alveolar type II epithelial cells |
| CN116814533B (en) * | 2023-08-28 | 2023-11-03 | 苏州冠佲生物医药科技有限公司 | Cell culture medium for in vitro maintenance of phenotype and function of alveolar type II epithelial cells |
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