[Opening: Let's talk about the 'Extreme Challenge of the Cell World']
Does the SP2/0 cell cloning experiment sound like a heavenly book? To put it simply: take out a cell and let it split into a whole group of identical offspring.
It's like letting a person live alone on a deserted island and still have to reproduce a kingdom. Cells are the same - they are extremely sensitive to the environment, nutrition, temperature, cleanliness, and operating methods... If there is a slight mistake, they will "strike" or even "die for you to see".
So, the cloning rate of this experiment directly reflects the true ability of serum to nourish cells. And there are only a few laboratories in the country that can achieve stability, precision, and reproducibility in this matter.
We are one of them.
Why can we do it? Two trump cards, one is indispensable
The first card: Hardware "ceiling"
The experiment was not conducted on a regular countertop, but in a positive pressure clean area+C/D grade GMP clean environment. What does this mean? The dust particles and microorganisms in the air are strictly filtered, and the air that cells breathe inside is cleaner than that in the ICU.
We are also equipped with a high-end cell counter - counting cells does not require visual estimation, the machine reads accurately, and the error is minimized; High definition microscope - whether cells are growing well or not, whether there is contamination, at a glance, even the morphology of cell colonies can be captured clearly (see Figure 3).
One sentence: The environment is a 'sterile ward', and the equipment is a 'sharp eye'.
Second card: Software "textbook level"
No matter how good the hardware is, people have to keep up. Our experimenters have undergone rigorous training and are skilled in dilution, plating, fluid replacement, and counting. Each step is carried out in accordance with the standard SOP of the Veterinary Pharmacopoeia, with errors controlled within a very small range.
More importantly, we have established a mature quality control system: every batch of experiments, from cell recovery to clone counting, is fully recorded and traceable. On any given day, which operator used which bottle of serum and which holes were photographed with a microscope can be clearly identified.
The result is accurate and reliable, and the data is traceable throughout the entire process - this is the confidence of "compliance".
Not only showcasing skills, but also providing customers with a 'reassurance pill'
As upstream partners of animal protection vaccine companies and research laboratories, we deeply understand that serum quality directly determines the success or failure of vaccine research and scientific experiments.
The SP2/0 cell cloning experiment is often an extremely difficult "internal threshold" to overcome. In order to ensure that our serum and culture medium have passed the most rigorous tests before delivery to customers, we have taken the lead in turning this challenge into an internal 'standard validation action'.
We take this verification to the extreme just to achieve:
Stable and controllable process: Each batch of experiments has internal quality control controls to ensure minimal fluctuations in the performance of each serum bottle leaving the factory;
Accurate and reliable results: The extremely high cloning rate data proves that our product can truly support high standards of research and production;
Full traceability of data: access original records at any time, win absolute trust from customers through transparent research and development processes.
Logic remains unchanged: Our goal is not to provide testing reports to customers, but to deliver the highest quality products to them.
Conclusion: Some thresholds, crossing them is like a moat
The SP2/0 cell cloning experiment cannot be done in any laboratory. It tests not only equipment, but also people's understanding of cells, reverence for standards, and authenticity of data.
We are willing to showcase these 'invisible skills' because we believe that true strength is not afraid of being watched. We first "train" our products to the extreme in our own hands, and then hand them over to customers. This is the greatest responsibility for product quality.
If you are looking for serum or culture media with excellent performance and stable quality for vaccine development or scientific research experiments, you may want to take a look at our "pre validation" results. Let's use a perfect experimental data to prove what it means to 'entrust professional matters to professional people'.
Attachment: SP2/0 Cell Cloning Experiment Standard Procedure (Veterinary Pharmacopoeia Edition)
Words are unfounded, the experimental plan is presented. The following is the complete operation process that we strictly follow according to the Chinese Veterinary Pharmacopoeia. We welcome colleagues to exchange and correct us.
1、 Experimental principle
Cell cloning refers to a population of cells formed by the proliferation of individual cells through mitosis, with identical genetic material. The higher the cloning rate, the stronger the serum's ability to nourish cells.
2、 Experimental route
Cell recovery → continuous passage (1:3) → cell dilution → 96 well plate gradient plating → 48 hour culture → clone counting → clone rate calculation → result determination
3、 Experimental steps
1. Preparation of culture medium
Take a 15mL centrifuge tube and prepare MEM complete culture medium containing 20% of the tested serum/control serum; Filter and sterilize with a 0.22 μ m sterile filter membrane, store in a sealed container at 4 ℃, and use up within one week (the fresher the better).
2. Cell preparation
Select SP2/0 cells in good condition and in logarithmic growth phase; Discard the original culture medium, gently resuspend with serum-free MEM, and blow evenly; Cell count and adjust the concentration of the cell suspension to 300000 to 500000 cells/mL.
3. Cell plating (96 well plate)
(1) First, add serum-free MEM to a 96 well plate at a concentration of 100 μ L per well (without adding it to the first and seventh rows);
(2) Dilute the cell suspension with serum-free MEM 1:50 and add it to the first and seventh rows at a concentration of 200 μ L/well;
(3) Perform gradient dilutions of 1:100, 1:200, 1:400, 1:800, and 1:1600 in rows 2-6 and 8-12, respectively, with 8 wells at each dilution level, resulting in a final volume of 100 μ L per well;
(4) Add 100 μ L of MEM containing 20% serum to each well (from high dilution to low dilution);
(5) Gently shake well to avoid cell clustering and ensure single-cell plating.
4. Cloning formation
Put it into a 37 ℃, 5% CO ₂ incubator; Let it stand and cultivate for 48 hours, avoiding movement or shaking as much as possible to ensure neat growth of clones.
5. Clone Count
Count only 30-50 clones/wells of dilution in duplicate wells, and finally calculate the clone formation rate.
Calculation formula:
Absolute clone formation rate (%)=(total number of clones at this dilution ÷ total number of cells inoculated at this gradient) × 100%
Relative clone formation rate (%)=(clone rate of test serum ÷ clone rate of control serum) × 100%
4、 Laboratory Safety Guidelines (Must Read)
Complete ultra clean bench operation, wiped with 75% alcohol and disinfected with UV for 30 minutes after completion;
Antifreeze gloves, goggles, and explosion-proof pipes must be worn during liquid nitrogen operations;
Alcohol and trypsin should be kept away from open flames, and any leaks should be dealt with promptly;
After high-pressure sterilization at 121 ℃ for 30 minutes, the experimental waste liquid is treated as medical waste;
Immediately isolate the sample upon discovering contamination, thoroughly disinfect it, and report it;
Standardize the use of instruments, promptly handle emergencies and report them.
5、 Experimental environment guarantee
This experiment relies on high standard hardware configurations such as positive pressure clean area and C/D GMP clean environment, equipped with high-end cell counter, high-definition microscope and other precision equipment, fully meeting the environmental requirements of the Veterinary Pharmacopoeia for sterile experiments. By cooperating with mature quality control systems and standardized operating procedures, we aim to achieve stable and controllable experimental processes, accurate and reliable results, and full traceability of data, providing compliant, efficient, and reliable quality verification and research and development support services for vaccine and biopharmaceutical enterprises.
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