Selection of aptamers for mature white adipocytes by cell SELEX using flow cytometry.
BACKGROUND
Adipose tissue, mainly composed of adipocytes, plays an important role in metabolism by regulating energy homeostasis. Obesity is primarily caused by an abundance of adipose tissue. Therefore, specific targeting of adipose tissue is critical during the treatment of obesity, and plays a major role in overcoming it. However, the knowledge of cell-surface markers specific to adipocytes is limited.


RESULTS
We applied the CELL SELEX (Systematic Evolution of Ligands by EXponential enrichment) method using flow cytometry to isolate molecular probes for specific recognition of adipocytes. The aptamer library, a mixture of FITC-tagged single-stranded random DNAs, is used as a source for acquiring molecular probes. With the increasing number of selection cycles, there was a steady increase in the fluorescence intensity toward mature adipocytes. Through 12 rounds of SELEX, enriched aptamers showing specific recognition toward mature 3T3-L1 adipocyte cells were isolated. Among these, two aptamers (MA-33 and 91) were able to selectively bind to mature adipocytes with an equilibrium dissociation constant (Kd) in the nanomolar range. These aptamers did not bind to preadipocytes or other cell lines (such as HeLa, HEK-293, or C2C12 cells). Additionally, it was confirmed that MA-33 and 91 can distinguish between mature primary white and primary brown adipocytes.


CONCLUSIONS
These selected aptamers have the potential to be applied as markers for detecting mature white adipocytes and monitoring adipogenesis, and could emerge as an important tool in the treatment of obesity.
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea.
Eun Young
EY
Kim
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea; Department of Functional Genomics, University of Science and Technology (UST) of Korea, Daejeon, Republic of Korea.
Ji Won
JW
Kim
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea.
Won Kon
WK
Kim
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea.
Baek Soo
BS
Han
Medical Proteomics Research Center, KRIBB, Daejeon, Republic of Korea.
Sung Goo
SG
Park
BioNanotechnology Research Center, Bioconvergence Research Institute, KRIBB, Daejeon, Republic of Korea.
Bong Hyun
BH
Chung
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea; Department of Functional Genomics, University of Science and Technology (UST) of Korea, Daejeon, Republic of Korea.
Sang Chul
SC
Lee
Research Center for Integrated Cellulomics, KRIBB, Daejeon, Republic of Korea; Department of Functional Genomics, University of Science and Technology (UST) of Korea, Daejeon, Republic of Korea.
Kwang-Hee
KH
Bae
D052437:Adipocytes, Brown / Q000378:metabolism; D052438:Adipocytes, White / Q000378:metabolism*; D000818:Animals; D052157:Aptamers, Nucleotide / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D005434:Flow Cytometry; D015723:Gene Library; D006801:Humans; D051379:Mice; D009928:Organ Specificity / Q000235:genetics; D052156:SELEX Aptamer Technique
D052157:Aptamers, Nucleotide