Chemical Cytoprotection for Stress-Free Use of Human Stem Cells in Basic and Translational Research

Dr. Ilyas Singec, Director, Stem Cell Translation Laboratory (SCTL), NIH Regenerative Medicine Program
What You Will Learn:
  • Quick and easy methods for isolating single cells for downstream applications
  • How to enhance cell survival of genetically stable human pluripotent stem cells (hPSCs)
  • How to ensure safe and robust stem cell growth with a novel, four-part small molecule cocktail
Record Date: November 9, 2021
Clinical translation of human pluripotent stem cells (hPSCs) requires advanced strategies that ensure safe and robust long-term growth and functional differentiation. We recently reported the development of a new small molecule cocktail that can be used as a universal end-to-end solution for stress-free culture of human cells. This four-part small molecule cocktail, which we named CEPT (Chroman 1, Emricasan, trans-ISRIB, Polyamines), enhanced cell survival of genetically stable hPSCs by simultaneously blocking several stress mechanisms that otherwise compromise cell structure and function. CEPT improved routine cell passaging, cryopreservation/thawing of pluripotent and differentiated cells, embryoid body and organoid formation, single-cell cloning, genome editing, and new iPSC line generation. Thus, CEPT represents a unique poly-pharmacological strategy for comprehensive cytoprotection, providing a new rationale for efficient and safe utilization of hPSCs. Conferring cell fitness by multi-target drug combinations may become a common approach in cryobiology, drug development, and regenerative medicine.

Fast, Gentle and Easy Single Cell Isolation

Mason Smith, Ph.D., Sr. Applications Scientist, Namocell
What You Will Learn:
  • Latest advancements in single cell sorting and dispensing technology
  • Discuss common pitfalls in several single cell workflows and practical tips to optimize results
  • Learn about novel single cell applications using innovative single cell sorting and dispensing technology
Record Date: October 7, 2021
Single cell sorting and isolation has traditionally been a bottleneck in the workflows for single cell cloning, single cell genomics, rare cell isolation, synthetic biology and other applications. Existing techniques such as fluorescence-activated cell sorting (FACS) often leads to compromised cell viability and poor outgrowth, contamination, clogging, in addition to the operational complexity and high maintenance requirement, while manual limiting dilution is inefficient, unreliable and highly labor-intensive. As such, there has been an urgent need for fast and easy single cell dispensing technology that is at the same time gentle to the cells and can preserve their viability and integrity. There is also a need for quick sorting of tens of thousands of cells for single cell genomics, such as sample prep for 10x Genomics workflow.

In this presentation, the speaker will discuss an innovative technology to address these needs. It combines the benefits of three key technologies: microfluidics, flow cytometry and liquid dispensing. This will simplify and empower a number of single cell applications, including cell line development, single cell genomics and proteomics, cell line engineering and CRISPR, monoclonal antibody development, synthetic biology, and rare cell isolation such as circulating tumor cells (CTCs) and circulating fetal cells.

Utilizing RNA and DNA Plate-Seq Workflows for Low-Input or Rare Single-Cell Genomics

Ioanna Andreou, Ph.D., Associate Director NGS & Applications Development, Qiagen
Felix Alonso-Valenteen, Ph.D., Field Application Scientist, Namocell
Applications: Single Cell Genomics
What You Will Learn:
  • Considerations, challenges and new technologies for low-input or rare single-cell genomics
  • Latest tips for low-input NGS
Record Date: April 26, 2022
Single-cell studies in recent years have revealed that organisms are composed of thousands of unique and unrepeatable cell types. Conducting single-cell genomic analyses using next generation sequencing (NGS) methods, similar to low-input sequencing, has traditionally been challenging due to the limited amount of genomic DNA present in single cells. This limitation becomes even more pronounced when the studied population is small, such as in the case of rare cells. High-fidelity (HiFi) and high-quality genomic DNA amplification is essential for single-cell sequencing, which is strongly dependent on the quality of the isolated cells. Therefore, the method used to isolate single cells becomes an important consideration for ensuring cell viability and nucleic acid integrity. Namocell’s patented gentle sorting and cell enrichment technology overcomes the aforementioned challenges to provide a higher quantity and quality of cell samples for sequencing. As a result, a complete and accurate picture of the studied samples can be achieved.