Stable Cell Lines Construction Service
Alpha Lifetech has been deeply engaged in the field of stable cell line construction process development for many years. It has built a perfect stable cell line construction process platform, with more than 300 kinds of tumor cells and primary cells, that can deliver stable cell lines while saving time for scientific research or project research, and facilitating subsequent production. Alpha Lifetech can also provide customers with services such as strain process development, strain screening, strain construction, recombinant protein, and antibody production, providing a research basis for the new drugs.
Alpha Lifetech has a lentivirus-infected cell platform, which can customize the experimental process for different types of cell stable transmutation strains. We used gene knockout technology to construct CRISPR/Cas9 knockout cell lines. Gene overexpression and gene silencing techniques were used to stabilize transmutation strains. Gene site-specific integration completes vertex integration cell line construction services.
Introduction to stable cell line construction
A stable cell line refers to the cell line that integrates foreign genes into the host cell genome so that foreign genes can be stably expressed in the host cell for a long time. The principle is to reduce the foreign gene to a vector with a certain resistance, transfect host cells, integrate the foreign gene into the host chromosome, and screen the resistance genes contained in the vector to obtain stable cell lines with high expression of the target protein or stable silencing of the target gene that meet the requirements of the experiment.
Alpha Lifetech has an independent storage workshop, the perfect cell freezing and storage management system, and more than 300 kinds of tumor cells and primary cells for you to choose from, dedicated to your scientific research.
Stable cell line construction process

Method of stable cell line construction
The stable cell line construction methods include lentivirus-mediated, transposon-mediated, CRISPR/Cas9-mediated, and plasmid screening.
lentivirus-mediated is one of the most commonly used methods to construct stable cell lines. This approach has the advantage of infecting almost all types of cells and integrating genetic material into the host cell genome for stable expression over a long period. However, this system has certain limitations, and lentivirus has a limited packaging capacity, so it is not suitable for transcription of genes with long transcription regions
Compared with lentivirus-mediated, transposon-mediated has no restriction on gene size, and with the continuous optimization of technology, the efficiency of transposon insertion has been improved several times. The disadvantage is that certain restrictions on primary cells and cells are difficult to transfect.
CRISPR/Cas9-mediated contains crispr gene knockout and crispr gene knockin. Crispr gene knockout refers to introducing DNA double-strand severing, resulting in a loss of function of the target gene. Crispr gene knockin refers to the foreign DNA fragments into specific genes, which can be used to repair defective genes or introduce new functional genes.
Plasmid screening is the most primitive cell line construction scheme, which only needs to successfully transfect the target plasmid into the cell and use the corresponding resistance in long-term screening. Then a certain probability of obtaining stable cell lines.
Methods of constructing different stable cell lines
Mediating Method | Lentivirus System | Transposon System | CRISP/Cas9-mediated | Plasmid Screening |
---|---|---|---|---|
Carrier Capacity | 5 kb | 10 kb | 5 kb | 10 kb |
Mediated Efficiency | High | Normal |
Normal | Low |
Timeline | 1-2 weeks | 3-4 weeks | 3-4 weeks | 4-6 weeks |
Advantage | Fast and efficient | Security and stability | Fixed point integration | Lower cost |
Shortcoming | Biosafety risk |
Expression abundance
normal
|
CAS9 transduction is difficult and costly | Low efficiency and instability |
Technical difficulties in the construction of stable cell lines
Before scaling up the process, a small bioreactor is used to determine parameters such as gas, culture fluid system, minimum hydrodynamic stress applied to the cells, etc., to ensure cross-scale culture. However, even if the corresponding process parameters are determined, the environment of large bioreactors in the producer will be uneven, and it is easy to fail. Alpha Lifetech has many years of experience in process scale-up culture, which can avoid mistakes to a certain extent, reduce risks, and greatly improve the success rate. At the same time, Alpha Lifetech ensures the consistency of raw materials and media batches used, reducing the failure rate of cell culture process scale-up.
Alpha Lifetech has a comprehensive stable cell line process development platform to ensure the stability of cell lines during production. We can also provide you with customized services to meet your needs.
Stable Cell Lines Construction Service content
Content | Timeline |
Culture HEK293 cell line provided by the customer. | 3-5 months |
Construction of sgRNA-Cas9 plasmid (with screening gene G418). | |
The sgRNA-Cas9 and repair DNA template were co-transferred to the HEK293 cell line. | |
Monoclonal cell line screening (GFP gene knockin positive clone). |
|
Delivery: Vector, GFP gene knock-in strain, sequencing results, all raw data, lab report |

Cells can be traced
The cells are purchased from professional organizations such as ATCC and can provide proof to meet your registration requirements Needs.

Production advantage
Alpha Lifetech has many years of experience to meet diverse development needs; We have a variety of cells, an independent storage workshop, perfect cell freezing, and a storage management system.

project Quality control
Each project undergoes strict quality control processes to ensure the delivery of cell lines that meet high standards to customers.

Technical advantage
In addition to engineers with extensive cellular experience, we have advanced technology evolutionary algorithms to improve reactor conditions and modify process parameters.
If you have any questions, please feel free to contact us at any time.
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