Cell Immortalization Service

Immortalized cell lines are cells that have been manipulated to proliferate indefinitely and can thus be cultured for long periods of time. They are either tumorous cells that do not stop dividing or cells that have been artificially manipulated to proliferate indefinitely and can, thus, be cultured over several generations to meet specific needs. KMD Bioscience has extensive research experience in cellular immortalization. We have already successfully constructed the immortalized cell lines derived from human, mouse and rat. Based on the dedicated scientific teams and advanced experimental cell immortalization platforms, KMD Bioscience has developed an efficient virus transfection technology. We offer a comprehensive range of reagents and engineered cell lines to meet your research needs. If you have any questions about production, please contact us. Our scientists will provide a reliable and risk-free cellular immortalization service to save your precious time and energy. Scientists at KMD Bioscience have focused on the development of immortalized cell line and have had many successful cases. We will escort your scientific research work!

Necessity of Cell Immortalization:

Although the primary cells have similar chromosome numbers as parent tissues and have specialized biochemical properties (growth factor and hormone secretion) as parent tissues, they have a limited lifespan/limited number of cell divisions, considerable variations in population and preparation, and are more susceptible to contamination. Cell immortalization could overcome those problems. Compared to primary cells, they tend to grow more quickly and can grow up to higher cell density, they are a uniform cell type, and most donor sources are the same therefore the batch-to-batch variation is less and they also maintain most cellular characteristics of the parent cells.  Using immortalized cell lines always saves money and time for long-term projects.

Fig 1. Cell immortalization procedure

Development of immortalized cell line:

The development of immortalized cell line involves transforming primary cells, which have a limited lifespan, into cells that can proliferate indefinitely under laboratory conditions. The following are the steps and methods for development of immortalized cell line：

1. Selection of Primary Cells

The first step is choosing the primary cells to be immortalized. KMD Bioscience can provide a variety of primary cells including fibroblast cell line, cardiomyocytes or hepatocyte cell line，or customers can also provide their own primary cells for research purposes.

2. Methods of cell Immortalization

--SV40 (Simian Virus 40) immortalization system

SV40 immortalization refers to the process of using Simian Virus 40 (SV40) genetic elements to extend the lifespan of cells in vitro, effectively making them immortal. The key to SV40 immortalization ability lies in the viral large T antigen (Tag), a protein that can disrupt normal cell cycle control and counteract cellular senescence mechanisms, leading to unlimited cell division. SV40 T antigen immortalization has facilitated the development of numerous cell lines used in diverse areas of biomedical research, from cancer biology to genetic studies, providing invaluable insights into cellular processes and disease mechanisms.

-- The human telomerase reverse transcriptase (hTERT) cell immortalization system

hTERT immortalization refers to a method used to extend the lifespan of primary cells through the ectopic expression of the human telomerase reverse transcriptase (hTERT) gene. Telomerase is an enzyme that adds telomeric DNA to the ends of chromosomes, countering the natural shortening that occurs with each cell division and which ultimately leads to cellular senescence. By expressing hTERT, cells can maintain their telomeres over many divisions, effectively bypassing the senescence barrier and achieving a state of immortality while retaining many of their primary characteristics.

-- Human papillomavirus (HPV) E6/E7 cell immortalization system

-- Epstein-Barr virus (EBV) cell immortalization system

-- MycT58A cell immortalization system

-- RasV12 cell immortalization system

3. Transfection and Selection

The chosen method for immortalization is introduced into the primary cells via transfection (for DNA-based methods) or infection (for viral vectors). Following this, cells must be selected for successful immortalization, often through antibiotic resistance markers included in the vector or by identifying cells that continue to proliferate beyond the expected lifespan of primary cells.

4. Verification of Immortalization

Cells believed to be immortalized are then subjected to tests to confirm their continuous growth potential. This includes:

- -Growth Curve Analysis: Assessing the ability of the cell line to proliferate over an extended period.

-- Telomerase Activity: For cell lines immortalized through telomerase activation, verifying the presence of telomerase activity.

-- Karyotyping: Analyzing the chromosomes for abnormalities that may indicate successful immortalization or unwanted mutations.

5. Characterization

--Morphological Assessment: Observing cell shape and growth patterns.

--Genetic and Phenotypic Stability: Ensuring the cell line maintains stable characteristics over passages.

-- Functional Tests: Assessing the cell line's ability to perform specific functions relevant to its intended use, such as expressing particular proteins or responding to stimuli.

Cell immortalization Service Content:

Customer Provides For Cell Immortalization:

-- The Target Cells

-- At least 4 tubes (＞1 X 106 cells/tube and dry ice transport)

-- Culture conditions

If your cells require media other than DMEM and RPMI, we will require 1-2L of media and any applicable growth factors that you provide for the project.

KMD Bioscience Deliverables:

-- 2 tubes (＞1 X 106 cells/tube) immortalized cells

-- Experimental report: morphological assessment; q-PCR analysis; Microbial contamination test results

Cell immortalization Service Highlights:

-- Custom service: provide different immortalization methods according to customer needs

-- PCR analysis of transgene expression in detail

-- Short time and high efficiency

-- Affordable price with the best quality

--Immortalization of B cells and immortalized macrophages cells

How to Order 

If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-400-621-6806;