Dr. Lin Wei, CEO and founder of inonecell, told arteria network that the first contact with single-cell sequencing was in 2014. At that time, bulk RNA sequencing was still the main means of transcriptome research in cell biopsy tissues, which released RNA from tissues and then carried out high-throughput quantitative detection of whole transcriptome.

In 2012, fluidigm, a listed company in the United States, launched C1 system, a smart SEQ cDNA library solution with 96 single cell chambers, which makes it possible to sequence and quantify the RNA of 96 cells. I have a strong intuition that for hundreds of millions of cells, it is limited to take only 96 sample cells for measurement to describe the characteristics of cells, but it is exciting enough to have fine detection. Once the cell flux increases to tens of thousands, the application scenario of single cell sequencing will have unlimited possibilities.

In 2016, 10x genomics launched the first single-cell high-throughput sequencing instrument chromium, which realized the construction of single-cell high-throughput cDNA library from the equipment level. Listed on Nasdaq, the company is currently the flagship company in the field of single-cell gene sequencing, with a market value of US $10 billion at the beginning of 2020. At the same time, Dr. Lin Wei’s laboratory also started the development of single cell sequencing project, and published related research papers based on the application of droplet high flux single cell technology framework with his collaborators.

Before the founding of Yinwan cell, Lin Wei was a doctor of computer biology at Tsinghua University, and later served as a postdoctoral fellow at the Cancer Institute of the National Institutes of health. He was a senior researcher at SAIC, a fortune 500 government consulting company, a senior bioinformatics expert at lengquangang functional genomics laboratory, and a Research Group Director (independent PI) at the Institute of immunology, Baylor medical center, Dallas, Texas. He has published many papers as a major participant in nature, science, cell and other top journals, and is an expert in single cell system development and application.

Another core technology member of the team, Dr. Wang Chunming, is a postdoctoral fellow of the Institute of immunology, Baylor medical center, Dr. Lin Wei. He has many years of research experience in the fields of biological signal detection, implementation of microfluidic chip technology, application and development of second-generation deep sequencing in biomedical field, etc. What’s more, many senior experts and engineers from Xi’an Institute of Optics and precision machinery, Chinese Academy of sciences joined the team. After just one year’s trial, Dr. Lin Wei’s team has achieved phased results in the research and development of single cell devices.

At the same time, the team also published papers on intelligent recognition of single-cell transcriptome, and launched the cancer single-cell mapping program. In June 2019, the team launched the first prototype of single-cell cDNA automatic library equipment with high-throughput and multi-channel detection, Dr. Lin Wei has established in-depth cooperation with many well-known scientific research and clinical institutions at home and abroad, including the city of hope Institute of translational genomics, Scripps Institute, Salk Institute, Renji Hospital of Shanghai Jiaotong University and Xiangya Hospital of Central South University.

In recent years, single cell sequencing and research topics have grown rapidly. Data show that in 2019, 1338 scientific papers have been published in PubMed, with a year-on-year increase of 102.4% compared with 661 in 2018. However, the prices of single-cell sequencing reagents, consumables and technical services remain high. Taking 10x genomics as an example, the service fee for data generation of a single-cell tissue sample is 20000-40000, which makes data analysis more difficult than data generation, and the service fee is also larger.

“If the data generation service fee is reduced to less than 10000, the single cell sequencing market will expand rapidly and reach the level that can be paid by the market. A powerful new detection technology is like a child getting a microscope. If it is not limited by the cost, he will put sand, leaves, insects and other things under the microscope to satisfy people’s strong curiosity. ” Dr. Lin Wei said, “at present, the industrial layout of domestic single cell sequencing enterprises is that a large number of third-party scientific research service providers purchase equipment, and then provide professional and complex single cell sequencing services to other scientific research institutions.

High throughput and multichannel single cell sequencing equipment will be applied in medical research

However, single-cell RNA sequencing has higher requirements for sample freshness. If it can be easily used by end users, it will be more ideal. This is also the direction we choose to cut into the industry. We start from the development and production of easy-to-operate integrated single-cell sequencing library detection equipment, give full play to the advantages of photoelectric measurement and control technology of the cooperative team, and produce equipment with superior performance and simple operation, And strive to reduce the cost of data generation and quickly establish market advantages. ” According to the introduction of Dr. Lin Wei, the reagent consumables cost of the new platform is less than 1 / 3 of the market benchmarking enterprises after the team’s whole machine optimization.

On the whole, single cell sequencing is to sequence the genetic material including DNA and RNA at the level of a single cell, and realize qualitative and quantitative analysis with a single cell as a unit, so as to identify the type and function of highly complex multicellular tissue and the changes and variations of specific cell health or state. However, the conventional detection based on the whole cell of the sample is based on the whole level of a large number of cells, and the genetic analysis of the whole tissue cell population is lost a lot of details.

From the perspective of operation process, the core of single cell sequencing technology needs a device to mark the cell identity of genetic material from different cells, and then conduct ngs deep sequencing on this basis.

Single cell preparation usually uses mechanical method, enzymatic method or other combination scheme to dissociate and digest tissue to form single cell suspension buffer. Single cell separation and DNA / cDNA capture labeling are the key steps of single cell sequencing, which are closely related to the quality and accuracy of the final single cell data. Among them, efficient microfluidic chip and precise photoelectric measurement and control are the key technologies of single cell separation and capture. In terms of core technology components of single cell sequencing equipment, Yinwan cell team has realized the independent production of most components.

Just as single cell sequencing is one of the application scenarios of NGS deep sequencing, it is a derivative application of ngs. As a kind of central node biotechnology, NGS has covered many application fields and derived various deep sequencing related application scenarios, such as genome, epigenetic group, microbiome, metagenome sequencing and so on. Dr. Lin Wei predicted that single cell sequencing itself will become another new central node of biotechnology, such as ngs deep sequencing technology leading a new industry. He hopes that the new single cell device developed by Yinwan cell will play an important role in the detection and application scenarios with multicellular tissue as the research object, just like Illumina’s ngs sequencing device.

It has been revealed that Yinwan cell has established cooperation with a number of domestic and foreign well-known scientific research and medical institutions to launch equipment, carry out scientific research cooperation activities, synchronously test and optimize equipment and technology, and sell reagents and consumables. The goal of scientific research cooperation is to jointly publish academic papers, rather than just provide equipment, reagents, consumables, single cell data generation or credit analysis services. To make a comprehensive market layout for the upstream and downstream technology application of single cell sequencing.

     

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