Method to purify and analyze heterogeneous senescent cell populations using a microfluidic filter with uniform...

Area of Science:

• Biotechnology and Biomedical Engineering
• Cell Biology and Aging Research

Background:

• Senescent cells accumulate with age and contribute to age-related diseases.
• Current methods for isolating senescent cells often lack precision and can be time-consuming.
• Heterogeneity within senescent cell populations complicates understanding of aging mechanisms.

Purpose of the Study:

• To design and validate a novel diamond-structure (DS) microfluidic filter for precise purification of senescent cells.
• To investigate the heterogeneity of senescent cells based on size after separation.
• To explore the potential of size-based separation for studying aging, autophagy, and rejuvenation.

Main Methods:

• Fabrication and optimization of a diamond-structure (DS) microfluidic filter using simulation.
• Separation of senescent human dermal fibroblasts into size-based cohorts (large and small).
• Assessment of senescence markers, including lipofuscin accumulation and beta-galactosidase activity.

Main Results:

• The optimized DS microfluidic filter achieved a uniform flow field, enhancing the throughput of size-based separation.
• Separated senescent cells exhibited significant size distribution and phenotypic heterogeneity.
• Smaller senescent cells displayed a younger phenotype, while larger senescent cells showed an older phenotype.

Conclusions:

• Size-based separation using the novel DS microfluidic filter provides an effective method for purifying senescent cells.
• This approach enables a more precise investigation into the heterogeneity of aging phenotypes.
• The findings support the use of size-based subtyping for studying aging mechanisms, autophagy impairment, and rejuvenation strategies.