Directed Differentiation of Adipose-Derived Stem Cells Using Imprinted Cell-Like Topographies as a Growth Factor-Free Approach.

The influence of surface topography on stem cell behavior and differentiation has garnered significant attention in regenerative medicine and tissue engineering. The cell-imprinting method has been introduced as a promising approach to mimic the geometry and topography of cells. The cell-imprinted substrates are designed to replicate the topographies and dimensions of target cells, enabling tailored interactions that promote the differentiation of stem cells towards desired specialized cell types. In fact, by replicating the size and shape of cells, biomimetic substrates provide physical cues that profoundly impact stem cell differentiation. These cues play a pivotal role in directing cell morphology, cytoskeletal organization, and gene expression, ultimately influencing lineage commitment. The biomimetic substrates' ability to emulate the native cellular microenvironment supports the creation of platforms capable of steering stem cell fate with high precision. This review discusses the role of mechanical factors that impact stem cell fate. It also provides an overview of the design and fabrication principles of cell-imprinted substrates. Furthermore, the paper delves into the use of cell-imprinted polydimethylsiloxane (PDMS) substrates to direct adipose-derived stem cells (ADSCs) differentiation into a variety of specialized cells for tissue engineering and regenerative medicine applications. Additionally, the review discusses the limitations of cell-imprinted PDMS substrates and highlights the efforts made to overcome these limitations.

Regulation of cell fate by cell imprinting approach in vitro.

Cell culture-based technologies are widely utilized in various domains such as drug evaluation, toxicity assessment, vaccine and biopharmaceutical development, reproductive technology, and regenerative medicine. It has been demonstrated that pre-adsorption of extracellular matrix (ECM) proteins including collagen, laminin and fibronectin provide more degrees of support for cell adhesion. The purpose of cell imprinting is to imitate the natural topography of cell membranes by gels or polymers to create a reliable environment for the regulation of cell function. The results of recent studies show that cell imprinting is a tool to guide the behavior of cultured cells by controlling their adhesive interactions with surfaces. Therefore, in this review we aim to compare different cell cultures with the imprinting method and discuss different cell imprinting applications in regenerative medicine, personalized medicine, disease modeling, and cell therapy.

Precise Capture and Dynamic Release of Circulating Liver Cancer Cells with Dual-Histidine-Based Cell Imprinted Hydrogels.

Circulating tumor cells (CTCs) detection presents significant advantages in diagnosing liver cancer due to its noninvasiveness, real-time monitoring, and dynamic tracking. However, the clinical application of CTCs-based diagnosis is largely limited by the challenges of capturing low-abundance CTCs within a complex blood environment while ensuring them alive. Here, an ultrastrong ligand, l-histidine-l-histidine (HH), specifically targeting sialylated glycans on the surface of CTCs, is designed. Furthermore, HH is integrated into a cell-imprinted polymer, constructing a hydrogel with precise CTCs imprinting, high elasticity, satisfactory blood compatibility, and robust anti-interference capacities. These features endow the hydrogel with excellent capture efficiency (>95%) for CTCs in peripheral blood, as well as the ability to release CTCs controllably and alive. Clinical tests substantiate the accurate differentiation between liver cancer, cirrhosis, and healthy groups using this method. The remarkable diagnostic accuracy (94%), lossless release of CTCs, material reversibility, and cost-effectiveness ($6.68 per sample) make the HH-based hydrogel a potentially revolutionary technology for liver cancer diagnosis and single-cell analysis.

Macrophage cell morphology-imprinted substrates can modulate mesenchymal stem cell behaviors and macrophage M1/M2 polarization for wound healing applications.

Mesenchymal stem cells and macrophages (MQ) are two very important cells involved in the normal wound healing process. It is well understood that topological cues and mechanical factors can lead to different responses in stem cells and MQ by influencing their shape, cytoskeleton proliferation, migration, and differentiation, which play an essential role in the success or failure of biomaterial implantation and more importantly wound healing. On the other hand, the polarization of MQ from proinflammatory (M1) to prohealing (M2) phenotypes has a critical role in the acceleration of wound healing. In this study, the morphology of different MQ subtypes (M0, M1, and M2) was imprinted on a silicon surface (polydimethylsiloxane [PDMS]) to prepare a nano-topography cell-imprinted substrate with the ability to induce anti-inflammatory effects on the mouse adipose-derived stem cells (ADSCs) and RAW264.7 monocyte cell line (MO). The gene expression profiles and flow cytometry of MQ revealed that the cell shape microstructure promoted the MQ phenotypes according to the specific shape of each pattern. The ELISA results were in agreement with the gene expression profiles. The ADSCs on the patterned PDMS exhibited remarkably different shapes from no-patterned PDMS. The MOs grown on M2 morphological patterns showed a significant increase in expression and section of anti-inflammatory cytokine compared with M0 and M1 patterns. The ADSCs homing in niches heavily deformed the cytoskeletal, which is probably why the gene expression and phenotype unexpectedly changed. In conclusion, wound dressings with M2 cell morphology-induced surfaces are suggested as excellent anti-inflammatory and antiscarring dressings.

Towards Electrochemical Sensor Based on Molecularly Imprinted Polypyrrole for the Detection of Bacteria-Listeria monocytogenes.

Detecting bacteria-Listeria monocytogenes-is an essential healthcare and food industry issue. The objective of the current study was to apply platinum (Pt) and screen-printed carbon (SPCE) electrodes modified by molecularly imprinted polymer (MIP) in the design of an electrochemical sensor for the detection of Listeria monocytogenes. A sequence of potential pulses was used to perform the electrochemical deposition of the non-imprinted polypyrrole (NIP-Ppy) layer and Listeria monocytogenes-imprinted polypyrrole (MIP-Ppy) layer over SPCE and Pt electrodes. The bacteria were removed by incubating Ppy-modified electrodes in different extraction solutions (sulphuric acid, acetic acid, L-lysine, and trypsin) to determine the most efficient solution for extraction and to obtain a more sensitive and repeatable design of the sensor. The performance of MIP-Ppy- and NIP-Ppy-modified electrodes was evaluated by pulsed amperometric detection (PAD). According to the results of this research, it can be assumed that the most effective MIP-Ppy/SPCE sensor can be designed by removing bacteria with the proteolytic enzyme trypsin. The LOD and LOQ of the MIP-Ppy/SPCE were 70 CFU/mL and 210 CFU/mL, respectively, with a linear range from 300 to 6700 CFU/mL.

Engineered substrates incapable of induction of chondrogenic differentiation compared to the chondrocyte imprinted substrates.

It is well established that surface topography can affect cell functions. However, finding a reproducible and reliable method for regulating stem cell behavior is still under investigation. It has been shown that cell imprinted substrates contain micro- and nanoscale structures of the cell membrane that serve as hierarchical substrates, can successfully alter stem cell fate. This study investigated the effect of the overall cell shape by fabricating silicon wafers containing pit structure in the average size of spherical-like chondrocytes using photolithography technique. We also used chondrocyte cell line (C28/I2) with spindle-like shape to produce cell imprinted substrates. The effect of all substrates on the differentiation of adipose-derived mesenchymal stem cells (ADSCs) has been studied. The AFM and scanning electron microscopy images of the prepared substrates demonstrated that the desired shapes were successfully transferred to the substrates. Differentiation of ADSCs was investigated by immunostaining for mature chondrocyte marker, collagen II, and gene expression of collagen II, Sox9, and aggrecan markers. C28/I2 imprinted substrate could effectively enhanced chondrogenic differentiation compared to regular pit patterns on the wafer. It can be concluded that cell imprinted substrates can induce differentiation signals better than engineered lithographic substrates. The nanostructures on the cell-imprinted patterns play a crucial role in harnessing cell fate. Therefore, the patterns must include the nano-topographies to have reliable and reproducible engineered substrates.

Improving performance of cell imprinted PDMS by integrating boronate affinity and local post-imprinting modification for selective capture of circulating tumor cells from cancer patients.

Efficient capture of circulating tumor cells (CTCs) from cancer patients is an important technique that may promote early diagnosis and prognosis monitoring of cancer. However, the existing systems have certain disadvantages, such as poor selectivity, low capture efficiency, consumption of antibodies, and difficulty in release of CTCs for downstream analysis. Herein, we fabricated an innovative PEGylated boronate affinity cell imprinted polydimethylsiloxane (PBACIP) for highly efficient capture of CTCs from cancer patients. The antibody-free PBACIP possessed hierarchical structure of imprinted cavities, which were inlaid with boronic acid modified SiO2 nanoparticles (SiO2@BA), so it could specifically capture target CTCs from biological samples due to the synergistic effect of boronate affinity and cell imprinting. Furthermore, PEGylation was accurately completed in the non-imprinted region by the template cells occupying the imprinted cavity, which not only retained the microstructure of original imprinted cavities, but also endowed PBACIP with hydrophilicity. The artificial PBACIP could efficiently capture human breast-cancer cells from biological sample. When 5 to 500 SKBR3 cells were spiked in 1 mL mice lysed blood, the capture efficiency reached 86.7 ± 11.5% to 96.2 ± 2.3%. Most importantly, the PBACIP was successfully used to capture CTCs from blood of breast cancer patients, and the captured CTCs were released for subsequent gene mutation analysis. The PBACIP can efficiently capture and release CTCs for downstream analysis, which provides a universal strategy toward individualized anti-tumor comprehensive treatments and has great potential in the future cell-based clinical applications.

Specific and quantitative detection of bacteria based on surface cell imprinted SERS mapping platform.

Non-specificity and poor quantitative ability are the main challenges in surface-enhanced Raman scattering (SERS) technique, especially for the detection of bacteria in real samples. In this study, we presented a surface cell imprinted SERS mapping platform which is competent for the specific and quantitative detection of bacteria. The platform based on the fabrication of a surface cell imprinted substrate (SCIS) by which Escherichia coli (E. coli) can be captured and labelled by SERS tags which produces strong characteristic signal to indicate the capture of targets. We highlighted the specificity of this platform in the detection of E. coli, by comparing the performances toward Salmonella paratyphoid A, Bacillus subtilis, Enterococcus faecalis and Staphylococcus aureus. Upon integrating with SERS mapping technique, the platform displayed good quantitative ability toward E. coli with a wide linear range from 102 to 108 CFU/mL and a low detection limit of ∼1.35 CFU/mL. Moreover, this novel SERS analysis platform was proved to be effective for E. coli detection in real probiotic beverage and chicken breast meat samples. By fabricating different SCISs, this platform can be replicated for the detection of other bacteria, which provides a promising application for real sample testing.

The Combined Thermoresponsive Cell-Imprinted Substrate, Induced Differentiation, and "KLC Sheet" Formation.

Purpose: Stem cells can exhibit restorative effects with the commitment to functional cells.Cell-imprinted topographies provide adaptable templates and certain dimensions for thedifferentiation and bioactivity of stem cells. Cell sheet technology using the thermo-responsivepolymers detaches the "cell sheets" easier with less destructive effects on the extracellularmatrix (ECM). Here, we aim to dictate keratinocyte-like differentiation of mesenchymal stemcells (MSCs) by using combined cell imprinting and sheet technology. Methods: We developed the poly dimethyl siloxane (PDMS) substrate having keratinocytecell-imprinted topography grafted with the PNIPAAm polymer. Adipose tissue-derived MSCs(AT-MSCs) were cultured on PDMS substrate for 14 days and keratinocyte-like differentiationmonitored via the expression of involucrin, P63, and cytokeratin 14. Results: Data showed the efficiency of the current protocol in the fabrication of PDMSmolds. The culture of AT-MSCs induced typical keratinocyte morphology and up-regulatedthe expression of cytokeratin-14, Involucrin, and P63 compared to AT-MSCs cultured on theplastic surface (P < 0.05). Besides, KLC sheets were generated once slight changes occur in theenvironment temperature. Conclusion: These data showed the hypothesis that keratinocyte cell imprinted substrate canorient AT-MSCs toward KLCs by providing a specific niche and topography.

Imprinted Polydimethylsiloxane-Graphene Oxide Composite Receptor for the Biomimetic Thermal Sensing of Escherichia coli.

This work presents an imprinted polymer-based thermal biomimetic sensor for the detection of Escherichia coli. A novel and facile bacteria imprinting protocol for polydimethylsiloxane (PDMS) films was investigated, and these receptor layers were functionalized with graphene oxide (GO) in order to improve the overall sensitivity of the sensor. Upon the recognition and binding of the target to the densely imprinted polymers, a concentration-dependent measurable change in temperature was observed. The limit of detection attained for the sensor employing PDMS-GO imprints was 80 ± 10 CFU/mL, a full order lower than neat PDMS imprints (670 ± 140 CFU/mL), illustrating the beneficial effect of the dopant on the thermo-dynamical properties of the interfacial layer. A parallel benchmarking of the thermal sensor with a commercial impedance analyzer was performed in order to prove the possibility of using the developed PDMS-GO receptors with multiple readout platforms. Moreover, S. aureus, C. sakazakii and an additional E. coli strain were employed as analogue species for the assessment of the selectivity of the device. Finally, because of the potential that this biomimetic platform possesses as a low-cost, rapid, and on-site tool for monitoring E. coli contamination in food safety applications, spiked fruit juice was analyzed as a real sample. Reproducible and sensitive results fulfill the limit requirements of the applicable European microbiological regulation.

Highly Integrated Cell-Imprinted Biomimetic Interface for All-in-One Diagnosis of Heterogeneous Circulating Tumor Cells.

Single-cell capture and in situ analysis of circulating tumor cells (CTCs) in blood are of great significance for early cancer diagnosis, prognosis, and individualized treatment. However, designing an all-in-one platform that enables not only efficiently specific isolation of CTCs but also in situ analysis of heterogeneity and drug screening is challenging. Here, a cell-imprinted alginate hydrogel (CIAH) interface with all-in-one functions was developed for the capture, in situ analysis, and drug-response study at a single-cell level. Based on the equivalent morphology and "specific odor" left by template cells and supplemented by natural antibody, the CIAH interface exhibited outstanding performance in isolating CTCs from samples suffering from cancers. Beyond capture, the CIAH interface was also able to serve as a high-throughput platform for subpopulation analysis and drug response of heterogeneous CTCs. We demonstrated that the highly integrated multifunctional CIAH interface is a promising new tool for single-cell profiling of phenotypic heterogeneity and guiding of personalized anticancer therapy.

Visual detection of hepatocellular carcinoma cells with cell imprinted substrate and pH-sensitive allochroic-graphene oxide.

Herein, we integrate cell-imprinted substrate (CIS) and allochroic-graphene oxide (AGO) for specific visualization sorting of hepatocellular carcinoma cells. The state-of-the-art-of detection method relies on the enzyme linked immunosorbent assay (ELISA)-like sandwich strategy with hierarchical recognition. The target tumor cells are first selectively captured by the CIS based on cell imprinted recognition, and then specifically labeled with AGO by boronate affinity recognition between boronic acid on AGO and cis-diols on the surface of target cells. The selectively recognition of CIS for target template cells is verified by cell function experiments. It is also worth mentioning that the AGO can specifically recognize target tumor cells under physiological pH, and then perform signal amplification and output through pH-triggered allochroism. The CIS linked AGO for cell assay (CIS-AGO-CA) is successfully used for visualization detection of human hepatocarcinoma HLE cells from hepatocyte suspension. When the hepatocyte suspension is spiked with 1.0 × 105 cells, the recoveries of CIS-AGO-CA are 80.67 ± 4.33% for target HLE cells, and only 12.00 ± 1.00% for non-target Hep3B cells. It is worth emphasizing that the CIS-AGO-CA process is antibody-free. Therefore, this novel ELISA-like sandwich strategy is high specificity, cost-efficient and easy-to-use, and exhibits great prospect in the visualization sorting of tumor subpopulation.

Capture and Kill: Selective Eradication of Target Bacteria by a Flexible Bacteria-Imprinted Chip.

This paper reports an antibacterial chip that can selectively capture bacteria and kill them using low-voltage DC electricity. We prepared a bacteria-imprinted, flexible PDMS chip that can separate target bacteria from suspensions with high selectivity. The chip contained integrated electrodes that can kill the captured bacteria within 10 min by applying a low DC voltage. The used chip could be easily regenerated by solution immersion. Meanwhile, the PDMS chip showed good biocompatibility and inhibited adhesion of human blood cells. Our work points to a new strategy to address pathogenic bacterial contamination and infection.

Neural priming of adipose-derived stem cells by cell-imprinted substrates.

Cell-imprinting technology is a novel method for directing stem cell fate using substrates molded from target cells. Here, we fabricated and studied cell-imprinted substrates for neural priming in human adipose-derived stem cells in the absence of chemical cues. We molded polydimethylsiloxane silicone substrates on fixed differentiated neural progenitor cells (ReNcellTMVM). The ReNcellTMcell line consists of immortalized human neural progenitor cells that are capable to differentiate into neural cells. The fabricated cell-imprinted silicone substrates represent the geometrical micro- and nanotopology of the target cell morphology. During the molding procedure, no transfer of cellular proteins was detectable. In the first test with undifferentiated ReNcellTMVM cells, the cell-imprinted substrates could accelerate neural differentiation. With adipose-derived stem cells cultivated on the imprinted substrates, we observed modifications of cell morphology, shifting from spread to elongated shape. Both immunofluorescence and quantitative gene expression analysis showed upregulation of neural stem cell and early neuronal markers. Our study, for the first time, demonstrated the effectiveness of cell-imprinted substrates for neural priming of adipose-derived stem cells for regenerative medicine applications.

Molecularly imprinted polymers for the selective recognition of microorganisms.

Molecularly imprinted polymers (MIPs) emerged half a century ago have now attracted tremendous attention as artificial receptors or plastic antibodies. Although the preparation of MIPs targeting small molecules, peptides, or even proteins is straightforward and well-developed, the molecular imprinting of microorganisms still remains a big challenge. This review highlights the preparation of MIPs that reveal biomimetic specificity and selectivity towards microorganisms by creating the well-defined cell recognition sites. We present the state-of-the-art strategies for the expeditious synthesis of MIPs targeting microorganism including surface components imprinting, cell mediated lithography, and microcontact stamping. These receptor-like biomimetic materials have garnered increasing attention in different fields. In this review, we also describe the diverse applications of microorganism-imprinted polymers such as microbial activation, microbial fuel cells, and microorganism detection and sensing. The major challenges and further prospects on the design of microorganism-imprinted polymers is also outlined.

Effect of cell imprinting on viability and drug susceptibility of breast cancer cells to doxorubicin.

This study demonstrates the effect of substrate's geometrical cues on viability and the efficacy of an anti-cancer drug, doxorubicin (DOX), on breast cancer cells. It is hypothesized that the surface topographical properties can mediate the cellular drug intake. Pseudo-three dimensional (3D) platforms were fabricated using imprinting technique from polydimethylsiloxane (PDMS) and gelatin methacryloyl (GelMA) hydrogel to recapitulate topography of cells' membranes. The cells exhibited higher viability on the cell-imprinted platforms for both PDMS and GelMA materials compared to the plain/flat counterparts. For instance, MCF7 cells showed a higher metabolic activity (11.9%) on MCF7-imprinted PDMS substrate than plain PDMS. The increased metabolic activity for the imprinted GelMA was about 44.2% compared to plain hydrogel. The DOX response of cells was monitored for 24 h. Although imprinted substrates demonstrated enhanced biocompatibility, the cultured cells were more susceptible to the drug compared to the plain substrates. In particular, MCF7 cells on imprinted PDMS and GelMA substrates showed 37% and 50% higher in cell death compared to the corresponding plain PDMS and GelMA, respectively. Interestingly, the drug susceptibility of the cells on the imprinted hydrogel was about 70% higher than the cells cultured on imprinted PDMS substrates. Having MCF7 cell-imprinted substrates, DOX responses of two other breast cancer cell lines, SKBR3 and ZR-75-1, were also evaluated. The results support that cell membrane curvature developed by multiscale topography is able to mediate intracellular signaling and drug intake. STATEMENT OF SIGNIFICANCE: Research in biological sciences and drug discovery mostly rely on two dimensional (2D) cell culture techniques which cannot provide a reliable physiologically relevant environment. Lack of extracellular matrix and a large shift in physicochemical properties of conventional 2D substrates can induce aberrant cellular behaviors. While chemical composition, topographical, and mechanical properties of substrates have remarkable impacts on drug susceptibility, gene expression, and protein synthesis, the most cell culture plates are from rigid and plain substrates. A number of (bio)polymeric 3D-platforms have been introduced to resemble innate cell microenvironment. However, their intricate culture protocols restrain their applications in demanding high-throughput drug screening. To address the above concerns, in the present study, a hydrogel-based pseudo-3D substrate with imprinted cell features has been introduced.

Evaluation of Surface Structure of Escherichia coli Using Polypyrrole Matrix.

We propose a method to evaluate the surface structure of Escherichia coli focusing on the doping state of bacterial cells into polypyrrole (PPy) matrix. We found that the orientation of doping states of E. coli O rough was different from those of other serotypes of E. coli cells, which had O-antigen on their outer membrane. The results indicated that more than seventy percent of E. coli cells having O-antigen was horizontally doped into PPy matrix based on the chemical structure and the placement of O-antigen. On the other hand, the percentage for horizontal doping state of E. coli O rough cells was only approximately fifty percent. Moreover, the cells of each E. coli serotypes were specifically bound to their own shape-complementary cavities on the microspheres, but the binding affinity of E. coli O rough was a bit lower than that of other serotypes.

Cell shape affects nanoparticle uptake and toxicity: An overlooked factor at the nanobio interfaces.

HYPOTHESIS: It is now being increasingly accepted that cells in their native tissue show different morphologies than those grown on a culture plate. Culturing cells on the conventional two-dimensional (2D) culture plates does not closely resemble the in vivo three-dimensional (3D) structure of cells which in turn seems to affect cellular function. This is one of the reasons, among many others, that nanoparticles uptake and toxicology data from 2D culture plates and in vivo environments are not correlated with one another. In this study, we offer a novel platform technology for producing more in vivo-like models of in vitro cell culture. EXPERIMENTS: The normal fibroblast cells (HU02) were cultured on "pseudo-3D" substrates, made from cell imprinting approach. The respond of the cells to a model nanoparticle (gold nanorod) were compared in 2D and "pseudo-3D" cultures modes, by cytotoxicological assays. FINDINGS: It is illustrated here that the cells' respond to the exact same type of nanoparticles is majorly dependant in their shape. The use of "pseudo-3D" substrates which could partially mimic the shape of cells in vivo is strongly proposed as a means of better predicting the efficacy of the 2D cell culture plates.

Nanomedicine and advanced technologies for burns: Preventing infection and facilitating wound healing.

According to the latest report from the World Health Organization, an estimated 265,000 deaths still occur every year as a direct result of burn injuries. A widespread range of these deaths induced by burn wound happens in low- and middle-income countries, where survivors face a lifetime of morbidity. Most of the deaths occur due to infections when a high percentage of the external regions of the body area is affected. Microbial nutrient availability, skin barrier disruption, and vascular supply destruction in burn injuries as well as systemic immunosuppression are important parameters that cause burns to be susceptible to infections. Topical antimicrobials and dressings are generally employed to inhibit burn infections followed by a burn wound therapy, because systemic antibiotics have problems in reaching the infected site, coupled with increasing microbial drug resistance. Nanotechnology has provided a range of molecular designed nanostructures (NS) that can be used in both therapeutic and diagnostic applications in burns. These NSs can be divided into organic and non-organic (such as polymeric nanoparticles (NPs) and silver NPs, respectively), and many have been designed to display multifunctional activity. The present review covers the physiology of skin, burn classification, burn wound pathogenesis, animal models of burn wound infection, and various topical therapeutic approaches designed to combat infection and stimulate healing. These include biological based approaches (e.g. immune-based antimicrobial molecules, therapeutic microorganisms, antimicrobial agents, etc.), antimicrobial photo- and ultrasound-therapy, as well as nanotechnology-based wound healing approaches as a revolutionizing area. Thus, we focus on organic and non-organic NSs designed to deliver growth factors to burned skin, and scaffolds, dressings, etc. for exogenous stem cells to aid skin regeneration. Eventually, recent breakthroughs and technologies with substantial potentials in tissue regeneration and skin wound therapy (that are as the basis of burn wound therapies) are briefly taken into consideration including 3D-printing, cell-imprinted substrates, nano-architectured surfaces, and novel gene-editing tools such as CRISPR-Cas.

Near-Infrared Light-Responsive Hydrogel for Specific Recognition and Photothermal Site-Release of Circulating Tumor Cells.

Isolation of single circulating tumor cells (CTCs) from patients is a very challenging technique that may promote the process of individualized antitumor therapies. However, there exist few systems capable of highly efficient capture and release of single CTCs with high viability for downstream analysis and culture. Herein, we designed a near-infrared (NIR) light-responsive substrate for highly efficient immunocapture and biocompatible site-release of CTCs by a combination of the photothermal effect of gold nanorods (GNRs) and a thermoresponsive hydrogel. The substrate was fabricated by imprinting target cancer cells on a GNR-pre-embedded gelatin hydrogel. Micro/nanostructures generated by cell imprinting produce artificial receptors for cancer cells to improve capture efficiency. Temperature-responsive gelatin dissolves rapidly at 37 °C; this allows bulk recovery of captured CTCs at physiological temperature or site-specific release of single CTCs by NIR-mediated photothermal activation of embedded GNRs. Furthermore, the system has been applied to capture, individually release, and genetically analyze CTCs from the whole blood of cancer patients. The multifunctional NIR-responsive platform demonstrates excellent performance in capture and site-release of CTCs with high viability, which provides a robust and versatile means toward individualized antitumor therapies and also shows promising potential for dynamically manipulating cell-substrate interactions in vitro.