title: Method to purify and analyze heterogeneous senescent cell populations using a microfluidic filter with uniform fluidic profile.
abstract: To precisely purify and study aged (senescent) cells, we have designed, fabricated, and demonstrated a novel diamond-structure (DS) microfluidic filter. Nonuniform flow velocities within the microfilter channel can compromise microfluidic filter performance, but with this new diamond structure, further optimized via simulation, we achieve a uniform microfilter flow field, improving the throughput of size-based separation of senescent cells, as obtained by 39-passaged human dermal fibroblasts. After separating these aged cells into two groups, consisting of large- and small-sized cells, we assessed senescence by measuring lipofuscin accumulation and β-galactosidase activity. Our results reveal that even though these senescent cells had been equivalently passaged in culture, a high degree of size distribution and senescent phenotype heterogeneity was observed. In particular, the smaller-sized cells tended to express a younger phenotype while the larger aged cells demonstrated an older phenotype. We suggest that size-based separation of senescent cells, subtyped into small- and large-sized cohorts, offers an alternative method to purify such aged cells, thereby enabling more precise study of the mechanisms of aging, autophagy impairment, and rejuvenation.
authors: Well Aging Research Center, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyenggi-do, Korea.|Department of Biomedical Engineering, Konyang University , 158 Gwanjeodong-ro, Seo-gu, Daejeon, Korea.
authors: Minseok S
authors: MS
authors: Kim
authors: Well Aging Research Center, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyenggi-do, Korea.
authors: Seonghyeon
authors: S
authors: Jo
authors: Well Aging Research Center, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyenggi-do, Korea.
authors: Joon Tae
authors: JT
authors: Park
authors: Well Aging Research Center, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyenggi-do, Korea.
authors: Hyun Young
authors: HY
authors: Shin
authors: R&D Solution Laboratory, Samsung Electronics, Ltd., Maetan3-dong, Youngtong-gu, Suwon-si, Gyeonggi-do, Korea.
authors: Sun Soo
authors: SS
authors: Kim
authors: Open Innovation Group, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyeonggi-do, Korea.|Campus D, 20, Yangpyeong-ro 21-gil, Yeongdeungpo-gu, Seoul, Korea.|Samsung Advanced Institute of Health Sciences and Technology, Irwon-ro 81, Gangnam-gu, Seoul, Korea.
authors: Ogan
authors: O
authors: Gurel
authors: Well Aging Research Center, Samsung Advanced Institute of Technology, Samsung Electronics, Ltd., 130, Samsung-ro, Yeongtong-gu, Suwon-si, Gyenggi-do, Korea.
authors: Sang Chul
authors: SC
authors: Park
mesh_terms: D002469:Cell Separation / Q000295:instrumentation; D002478:Cells, Cultured; D016922:Cellular Senescence; D002648:Child; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D010316:Particle Size; D012867:Skin / Q000166:cytology; D013499:Surface Properties