(1401-B) Drastically Accelerated Directed Differentiation of Human Pluripotent Stem Cells into Melanocytes for Phenotypic Screening
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: Melanocytes are specialized cells primarily known for their role in producing melanin, the pigment responsible for the color of skin, hair, and eyes. Their functions and significance in the body extends beyond their contribution to coloration, and are multifaceted from protecting the skin from ultraviolet (UV) radiation to regulation of immune responses. Developing screening assays to understanding the complex roles of melanocytes opens avenues for research in dermatology, oncology, and neurobiology including diseases like vitiligo, where melanocytes are destroyed, or melanoma, a serious form of skin cancer. The differentiation of human pluripotent stem cells (hPSCs) into specific cell types is a cornerstone of regenerative medicine and disease modeling. However, the process is often slow and inefficient, particularly for the derivation of melanocytes where protracted protocols take over a month and several additional weeks of maturation are necessary before melanin production occurs. Here we demonstrate the rapid and efficient production of melanocytes from human induced pluripotent stem cells using directed differentiation in 9 days under defined media conditions. Macroscopic pigmentation is observed within an additional week of maturation. These melanocytes were characterized via immunocytochemistry and qPCR for the melanocyte specific markers SOX10, MITF, DCT, TYR, and TYRP1 and expression was compared to primary human melanocytes. To demonstrate their utility as a simple assay for screening purposes, large scale batches of melanocytes were manufactured and banked. Melanocytes were thawed into 96-well plates and after a week of maturation were used to screen and identify pigment regulating agents. We show that 4-butylresorcinol, an inhibitor of tyrosinase and tyrosinase-related protein-1, can macroscopically decrease pigmentation in only three days. This advance in melanocyte production holds substantial promise as a scalable, consistent tool for high-throughput phenotypic screenings, particularly in cosmetics and pigmentation disorders.