Professor Guo Guozhen after the 80s in the experiment

6 trillion tiny, complex cell structures make up the human body. Simply put, each of us is composed of a bunch of cells; every time we breathe, exercise, and think, the cells are working.

But people know little about cells.

Humans have taken a new big step - the famous academic journal "CELL" ("Cell"), which was launched yesterday, published the research results of the team of Professor Guo Guojun from the Stem Cell and Regenerative Medicine Center of Zhejiang University School of Medicine. Cost, high efficiency, fully localized high-throughput single-cell sequencing platform Microwell-seq, and used this platform to build the world's first mammalian cell map in a short time.

This will open a wonderful journey to bring humans into a fantasy world.

From "seeing" a group of cells

To a single cell

Cells are the smallest independent genetic unit of life. The living body begins to divide from a fertilized egg and gradually develops into an individual. The difference between the cells is getting bigger and bigger, and it bears different functions. Even if cells of the same function differ, an extreme example is that tumor cells are initially mutated by a normal cell.

In October 2017, American scientists and multinational scientists launched the "Human Cell Atlas Program", which envisages the classification and sequencing of all cells in the human body, systematically depicts the human cell map, and uses this key to deepen the diagnosis, monitoring, and treatment of diseases. Understanding.

Facebook founder Mark Zuckerberg funded the project and called it an international research project comparable to the Human Genome Project. In the past few years, single-cell sequencing has shown broad prospects in detecting tumor variability and prenatal diagnosis.

Traditional sequencing technology, "look" is a group of groups, cells, "read" is the average of a bunch of cellular genetic signals, so the specific performance of a single cell is easily overlooked. Single-cell sequencing, by "reading" the genetic information of a single cell, is a good response to the heterogeneity of the cell population.

For this reason, single-cell sequencing technology has become a hot spot for scientists in various countries, and further promotes human cognition of the mystery of life.

"Single-cell sequencing technology enables humans to more accurately resolve cell differentiation, regeneration, senescence, and pathology from the perspective of a single cell." Guo Guojun said, "This type of technology is bringing about a cell detection, classification, and identification. Methodological revolution."

Single cell sequencing cost

From one hundred yuan to two yuan

In the face of the ambition of the Cell Atlas program, technology has developed rapidly in recent years, and scientists from all over the world are competing to develop high-level single-cell sequencing platforms. One of the key questions is: How to quickly and efficiently detect a single cell in order to complete this huge amount of super "puzzles"?

Professor Guo Guojun's team introduced their solutions to the world in CELL: high-throughput, low-cost, simple-to-use, fully localized Microwell-seq single-cell sequencing platform. "We use microwell matrix, molecular labeling and amplification technology to achieve single-cell level analysis with high throughput and high precision, which solves the problem of too few single-cell nucleic acid substances in traditional sequencing, easy to lose, and high analysis cost."

Dr. Han Xiaoping (first author of the paper) showed the reporter a square piece with a side length of 3 cm - this is a microplate made of agarose. "There are 100,000 small pits with a diameter of 30 microns."

Scientists first use digestive enzymes to dissociate a relatively tight cell into a suspension of individual cells, and then dump it onto agarose microplates. "About 10,000 cells will fall into the pit with a radish." Medium.” This is the first step: capturing a single cell.

Thereafter, the scientists attached a "number" to each of the captured cells. Tens of thousands of magnetic beads with a diameter of 25 microns fall into the pit, sealing a single cell, while indexing the DNA. This is the second step, called the "magnetic bead index."

When preparing to index magnetic beads, the research team had searched all the biotech companies in China, but none of them could do it. Finally, the research team synthesized itself.

The third step is the conventional sequencing process, which resolves the expression profile of each cell. In the technique of Professor Guo Guojun's team, all mRNAs in a single cell, a genetic material capable of translating proteins, were measured.

"After analyzing a cell, it takes about 100 RMB, and with the platform we developed, it is less than 2 yuan, and the experimental operation is faster and easier." Guo Guozhen said that the Microwell-seq technology platform will promote the cutting-edge single-cell sequencing. The popularity and application of technology in basic scientific research and clinical diagnosis.

Draw the first

Mammalian cell map

Using the Microwell-seq technology platform, the team performed a systematic single-cell transcriptome analysis of more than 400,000 cells from nearly 50 organ tissues of different life stages of mice, and constructed the first mammalian cell map.

Guo Guojun called this map "cell map." The team described the major cell types of mice, histiocytic subtypes, stromal cell subtypes, vascular endothelial cell subtypes, and immune cell subtypes in each organ.

The study found that stromal cells from different tissues have completely different gene expression characteristics and exert important regulatory effects on the tissue-specific microenvironment. This suggests that scientists should not only consider tissue cells in tissue engineering and organ repair, but must also repair the corresponding stromal cell system.

"Human recognition of life is like exploring the vast universe. Single-cell sequencing gives us a more special 'observation' approach to exploring important substances not found in the 'Starry Sky' of life."

Guo Guozhen said. "Scientists can use this platform to conduct a mechanistic study of cell fate determination, which will have a profound impact on pre-transplant cell identification of regenerative medicine and cell-level diagnosis of clinical disease."

During the review process, the three reviewers of CELL consistently gave an “impressive” evaluation and believed that “the mouse map is bound to become a versatile biological data resource”.

The scientific community believes that the completion of the mouse cell map will also guide the construction of the next human cell map, and benefit from many fields such as cell biology, developmental biology, neurobiology, hematology and regenerative medicine.

"Our team has actually completed a vast project with one's own efforts. I hope that everyone has confidence in China's life science industry, technology and scientists. The Chinese are fully capable and technically able to draw a Chinese cell image."

Guo Guozhen said, "This single-cell sequencing technology is cost-effective, and all reagents and materials are completely domestically produced, and the commercialization prospects are very broad."

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