Construction of single-cell arrays and assay of cell drug resistance in an integrated microfluidic platform

Abstract

The cellular heterogeneity of tumors has played important roles in various tumor-related research areas and applications such as the cellular biology, metastasis and clinical diagnosis of tumors. Although several microfluidics-based single-cell separation and analysis techniques have been used in research into the cellular heterogeneity of tumors, further investigation is still required for studying the effect of the biomechanical (e.g., size and deformability) heterogeneity of cells on their biological characteristics (e.g., drug resistance and tumor-initiating features). Here, we established an integrated microfluidic platform for the construction of single-cell arrays and analysis of drug resistance. Using this device, high-throughput single-cell arrays could be easily obtained according to the biomechanical (size and deformability) heterogeneity of cells. To demonstrate the capability of the microfluidic platform, a proof-of-concept experiment was implemented by determining the vincristine resistance of single glioblastoma cells with different biomechanical properties. The results indicated that the biomechanics of tumor cells had significant implications for cell drug resistance; that is, small and/or more deformable tumor cells had higher drug resistance than large and/or less deformable tumor cells. This device provides a new approach for the isolation of single cells according to the different biomechanical properties of cells. Also, it possesses practical potential for studies of tumors on a single-cell level.

Application of microfluidic chip technology in pharmaceutical analysis: A review

“…It is composed of a culture medium reservoir, a syringe pump, an oxygenator, a bubble trap, and a chip and was imaged on a two-photon imaging system [114]; (E) Integrated microfluidic system for single-cell separation according to cell size and deformability, which combined multiobstacle architecture-like microstructural matrices and a microvalve system. Using this device, the biomechanical (size or deformability) heterogeneity of normal and induced glioblastoma cells was studied on a single-cell level [67].…”

“…An efficient droplet microfluidics-based approach assessed the dynamics of drug uptake, efflux and cytotoxicity in drug-sensitive and drug-resistant breast cancer cells [13]. Pang et al [67] established an integrated microfluidic platform for the construction of single-cell arrays and the analysis of drug resistance. Fig.…”

“…This review provides a new insight into the research of drug analysis. Table 1 describes the significant use of microfluidic chips in this area and summarizes the device features and related representative literature [7], [8], [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], [19], [20], [21], [22], [23], [24], [25], [26], [27], [28], [29], [30], [31], [32], [33], [34], [35], [36], [37], [38], [39], [40], [41], [42], [43], [44], [45], [46], [47], [48], [49], [50], [51], [52], [53], [54], [55], [56], [57], [58], [59], [60], [61], [62], [63], [64], [65], [66], [67].…”

A review on microwell and microfluidic geometric array fabrication techniques and its potential applications in cellular studies

“…The traps are designed to allow the smaller, more deformable cells to pass through holes and spacings between the traps, while larger, less deformable cells are caught in said traps. [ 250 ] Ultimately, precise control over the size and number of cells per trap can be achieved based on a uniquely designed pillar formation for any variety of targets using two‐photon‐lithography in controlled flow (Figure 17E). Although this requires sophisticated equipment, it is possible to isolate single or multiple particles with 100% trapping efficiency and eliminate the need for constant pressure to hold particles in place.…”

“…A, Stretchable asymmetry‐bracket‐shaped microtrap structure (SAMS) for tunable size traps, reprinted with permission from Xu et al [ 218 ] ; B, cell loading using deformation of cells in u‐shaped traps, reprinted, with permission, from Dura et al [ 248 ] ; C, cell sorting using bandpass filtering and cell isolation, reprinted, with permission, from Kim et al [ 249 ] ; D, application of biochemical cell heterogeneity for cell sorting and isolation, reprinted, with permission, from Pang et al [ 250 ] ; and E, real‐time trapping of particle using two‐photon‐lithography in a controlled flow, reprinted, with permission, from Xu et al [ 251 ] …”