Cell sampler units, cell sampler tools comprising the same and methods of cell sampling

Abstract

A cell sampling unit is disclosed, comprising: a plurality of micropillars for contacting a sample and acquiring cells from the surface of the sample by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the acquired cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end comprising a contact surface with frictional force; wherein each micropillar has a diameter of approximately 60-140 µm and a height of approximately 60-140 µm; wherein the plurality of micropillars are configured in an array arrangement; the micropillars are arranged at a rate of approximately 1000-1400 micropillars / cm 2 The density distribution between [specific parameters]. Other example embodiments are described herein. In some embodiments, the cell sampling unit is an easy-to-use, convenient, and reliable skin cell sampling method for male HPV screening.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202580006872.5 (22) Application Date 2025.01.13 (30) Priority Data 63 / 621,132 2024.01.16 US (85) PCT International Application Entering National Phase Date 2026.06.02 (86) PCT International Application Application Data PCT / CN2025 / 072058 2025.01.13 (87) PCT International Application Publication Data WO2025 / 152897 EN 2025.07.24 (71) Applicant: Chuangjian Medical (Hong Kong) Limited Address: Central, Hong Kong, China (72) Inventors: Gao Junxi, Huang Zhenhui, Zeng Kunyuan (74) Patent Agency: Shenzhen Yingchi Intellectual Property Agency Co., Ltd. 44658 Patent Attorneys Huang Xing'er and Wang Yijin (51) Int.Cl. C12M 1 / 26 (2006.01) A61B 10 / 02 (2006.01) C12Q 1 / 24 (2006.01) (54) Invention Title: Cell Sampler Unit, Cell Sampler Tool Containing the Same, and Cell Sampling Method (57) Abstract: A cell sampling unit is disclosed, comprising: a plurality of micropillars for contacting a sample and obtaining cells from the surface of the sample by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the obtained cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end, the free end comprising a contact surface with friction; wherein each micropillar has a diameter of approximately 60-140 µm and a height of approximately 60-140 µm. µm; wherein the plurality of micropillars are configured in an array arrangement; the micropillars are distributed at a density between approximately 1000-1400 micropillars / cm². Other exemplary embodiments are described herein. In some embodiments, the cell sampling unit is an easy-to-use, convenient, and reliable skin cell sampling unit for male HPV screening. Claims 2 pages Description 15 pages Drawings 35 pages CN 122555765 A 2026.08.11 CN 1 22 55 57 65 A 1. A cell sampling unit comprising: a plurality of micropillars for contacting a sample and acquiring cells from the surface of the sample by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the acquired cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end comprising a contact surface having a frictional force; wherein each micropillar has a diameter of approximately 60-140 µm and a height of approximately 60-140 µm;1. The plurality of micropillars are configured in an array arrangement; the micropillars are distributed at a density between approximately 1000-1400 micropillars / cm². 2. The cell sampling unit of claim 1, wherein the sample is derived from human skin, such as skin from the male genitalia for human papillomavirus (HPV) detection in men. 3. The cell sampling unit of claim 1, wherein the overall length of the cell sampling unit is approximately 1-2 cm and the width is approximately 1-3 cm. 4. The cell sampling unit of claim 1, wherein each micropillar is a cylinder, triangular prism, square prism, hexagonal prism, or pentagonal prism. 5. The cell sampling unit of claim 1, wherein each micropillar and / or the substrate is made of a biocompatible polymer material, such as polydimethylsiloxane (PDMS), thermoplastic polyurethane (TPU), and / or polycaprolactone (PCL). 6. A cell sampling unit for detecting human papillomavirus (HPV) in men, the cell sampling unit comprising a plurality of micropillars for contacting a sample and acquiring cells on the surface of the sample by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the acquired cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end comprising a contact surface with frictional force; wherein each micropillar is cylindrical with a diameter of approximately 60-140 µm and a height of approximately 100-140 µm; wherein the plurality of micropillars are configured in an array arrangement; the micropillars are distributed at a density of approximately 1000 micropillars / cm²; and wherein each micropillar and / or the base layer is made of polydimethylsiloxane (PDMS). 7. A cell sampling tool comprising: a cell sampling unit as claimed in any one of claims 1-6; and an actuator; wherein a base layer of the cell sampling unit is attached to or extends from the actuator, such that the cell sampling unit rubs against the sample under an external force applied to the actuator to obtain cells. 8. A cell sampling method comprising the steps of: rubbing a sample with a cell sampling unit as claimed in any one of claims 1-6 or a cell sampling tool as claimed in claim 7 to obtain cells therefrom. 9. The method of claim 8, wherein rubbing the sample comprises the steps of: placing the cell sampling unit or the cell sampling tool on a sample surface, the free end facing the sample surface, and rubbing the sample back and forth under an external force to collect the cells. 10. The method of claim 8 or claim 9, wherein the sample is human skin, such as male genital skin for human papillomavirus (HPV) detection in men. 11. The method of any one of claims 8-10, further comprising the steps of:Immerse the cell sampling unit or the cell sampling tool in the extraction solution to detach the cells from the sampling unit and to induce the cells to detach from the sampler under external force; and separate the detached cells from the extraction solution by centrifugal force, removing the supernatant to obtain a precipitate suitable for DNA extraction. 12. The method of any one of claims 8-10, further comprising the step of rubbing the sample, such as human skin, 1-20 times to obtain 5,000-50,000 cells or more. Claims 2 / 2 Page 3 CN 122555765 A Cell Sampler Unit, Cell Sampler Tool Including the Same, and Cell Sampling Method Cross Reference to Related Applications

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 621,132, filed January 16, 2024, pursuant to 35 USC § 119(e) (35 USC § 119(e)), the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to a cell sampler unit, a cell sampler tool including the same, and a cell sampling method. Background Art

[0003] Human papillomavirus (HPV) is the most common sexually transmitted infection (STI) in the world. Increasing evidence of a correlation between HPV and anogenital cancers has led to the development of HPV testing for both women and men. Cell sampling is a critical step in HPV testing because the viral load of HPV in infected tissue is typically very low. To date, different types of samplers have been developed for collecting HPV samples from the female cervix. However, no similar samplers have been developed for men. In many clinical settings, sandpaper and medical swabs are used together for sample collection. This two-step collection method has several drawbacks, such as causing discomfort and anxiety, and more importantly, a low success rate. Therefore, there is an urgent need to develop an easy-to-use genital cell sampler for male HPV DNA.

[0004] In some embodiments, a cell sampler unit, a cell sampler tool comprising the same, and a cell sampling method are provided. In some embodiments, a cell sampler with a novel microstructure is provided for skin cell sampling, such as male genital HPV sampling. In some embodiments, a "Bio-Sandpaper" (BioS) technology is provided, which mimics naturally occurring microstructures, such as bifunctional microstructures that mimic micron- to nanometer-scale bristles in organisms and the micro-patterned hairs on bee legs that capture pollen. Such biomimetic microstructures can easily extract and capture skin cells. To achieve this, in some embodiments, the microstructure contains numerous micropillars whose shape and size will be optimized to increase...The surface area to volume ratio of the biocompatible polymer surface is increased to generate high frictional force for skin cell collection. In some embodiments, the micropillar spacing and pattern are optimized to effectively capture and release skin cells. In some embodiments, the biocompatible polymer is tested to check the consistency of microstructure fabrication in the molding system, making the provided sampler feasible for future scale-up manufacturing. In some embodiments, the cell sampler prototype is made from a sampler head and operating rod with microstructures. In some embodiments, the provided cell sampler is an easy-to-use, convenient, and reliable skin cell sampler for male HPV screening.

[0005] Figure 1A is a photograph showing the mold design of a cell sampler unit with 42 x 35 micropillars according to an example embodiment.

[0006] Figure 1B is a schematic diagram showing the micropillar dimensions of a cell sampler unit with a height of 140 µm and a width of 140 µm according to an example embodiment. Instruction Manual 1 / 15 Page 4 CN 122555765 A

[0007] FIG1C is a photograph showing the main silicon wafer mold for manufacturing a cell sampler unit according to an exemplary embodiment.

[0008] FIG1D is a photograph showing a PDMS cell sampler unit of size 1 x 1.4 cm2 according to an exemplary embodiment.

[0009] FIG2A is a schematic diagram showing the steps of manufacturing a cell sampler head unit from PDMS according to an exemplary embodiment.

[0010] FIG2B is a photomicrograph showing a front view of a PDMS cell sampler head unit having a micropillar with a width of 140 µm under an optical microscope at 100x magnification according to an exemplary embodiment.

[0011] FIG2C is a photomicrograph showing a side view of a PDMS cell sampler head unit having a micropillar with a height of 140 µm under an optical microscope at 100x magnification according to an exemplary embodiment.

[0012] FIG3A is a schematic diagram showing the steps of manufacturing a cell sampler head from PCL according to an exemplary embodiment.

[0013] FIG3B is a photomicrograph showing a front view of a PCL cell sampler head unit having a micropillar with a width of 140 µm under a stereomicroscope at 80x magnification, according to an exemplary embodiment.

[0014] FIG3C is a photomicrograph showing a side view of a PCL cell sampler unit having a micropillar with a height of 108 µm under an optical microscope at 100x magnification, according to an exemplary embodiment.

[0015] FIG4A is a schematic diagram showing the steps of manufacturing a cell sampler head from TPU according to an exemplary embodiment.

[0016] FIG4B is a photomicrograph showing a front view of a TPU cell sampler head under an optical microscope at 100x magnification, according to an exemplary embodiment. The width of the micropillar is 140 µm.

[0017] FIG4C is a photomicrograph showing a side view of a TPU cell sampler head under an optical microscope at 100x magnification, according to an exemplary embodiment.Figure 5A is a schematic diagram showing the steps of analyzing the PDMS cell sampler head by scanning electron microscopy (SEM) according to an exemplary embodiment. Figure 5B is a SEM micrograph showing a front view of the PDMS cell sampler head at 100x magnification according to an exemplary embodiment. The sampler density is > 1000 micropillars / cm². Figure 5C is a side view of the PDMS cell sampler at 100x magnification according to an exemplary embodiment. The height and width of the micropillars are both 140 µm. Figure 6A is a schematic diagram (left) and a photograph (middle) of a silicon wafer mold having a cell sampler unit with 42 x 35 square micropillars, and a photograph (right) of a PDMS cell sampler head having a cell sampler unit with square micropillars, respectively, according to an exemplary embodiment.

[0022] Figure 6B is a photomicrograph showing the front view (left) and side view (right) of a PDMS sampler with square micropillars under an optical microscope at 100x magnification, according to an example embodiment. The height and width of the square micropillars are both 140 µm.

[0023] Figure 6C is a schematic diagram (left) and a photograph (middle) of a silicon wafer mold for a cell sampler unit with triangular micropillars, and a photograph (right) of the PDMS cell sampler unit with triangular micropillars, according to an example embodiment.

[0024] Figure 6D is a photomicrograph showing the front view (left) and side view (right) of a PDMS sampler with triangular micropillars under an optical microscope at 100x magnification, according to an example embodiment. The height and width of the triangular micropillars are both 140 µm.

[0025] Figure 7 is a graph showing the friction force of the cell sampler unit on synthetic skin, measured with reference to ASTM D1894, according to an example embodiment. In this figure, the COF of micropillars with different shapes was tested and compared.

[0026] Figure 8A is a schematic diagram showing the friction generated by the microstructure on synthetic skin, measured using a coefficient of friction (COF) tester according to an example embodiment. The figure shows different directions of movement of the triangular micropillars. The frictional force of the cell sampler head on the synthetic skin was measured with reference to ASTM D1894. The COF of the triangular micropillars in different directions of movement was tested and compared.

[0027] Figure 8B is a graph showing the static COF of the triangular micropillars in different directions of movement according to the example embodiment of Figure 8A. Figure 8C is a graph showing the dynamic COF of the triangular micropillars in different directions of movement according to the example embodiment of Figure 8A.

[0028] Figure 9A is a micrograph showing micropillars of various heights (60 µm, 100 µm, and 140 µm) prepared to optimize the micropillar height according to an example embodiment. (Refer to ASTM D1894)The frictional force of the cell sampler on synthetic skin was measured using ASTM D1894.

[0029] Figure 9B is a graph showing the static COF of micropillars of different heights according to the example embodiment of Figure 9A. The frictional force of the cell sampler on synthetic skin was measured with reference to ASTM D1894.

[0030] Figure 9C is a graph showing the dynamic COF of micropillars of different heights according to the example embodiment of Figure 9A.

[0031] Figure 10A is a photomicrograph of a cell sampler with micropillars of various widths (60 µm, 100 µm, and 140 µm) prepared according to the example embodiment for optimizing micropillar width, under an optical microscope at 100x magnification. The frictional force of the cell sampler on synthetic skin was measured with reference to ASTM D1894. The COF of samplers of different widths was tested and compared.

[0032] Figure 10B is a graph showing the static COF of micropillars of different widths according to the example embodiment of Figure 10A.

[0033] Figure 10C is a graph showing the dynamic COF of micropillars of different widths according to the example embodiment of Figure 10A.

[0034] Figure 11A is a photomicrograph under a 100x optical microscope showing cell samplers with various micropillar densities prepared according to the example embodiment for optimizing micropillar density. The dimensions of the cell samplers were observed under a 100x optical microscope. The friction of the cell samplers on synthetic skin was measured with reference to ASTM D1894. The COF of samplers with different densities was tested and compared.

[0035] Figure 11B is a graph showing the static COF of micropillars of different densities according to the example embodiment of Figure 11A.

[0036] Figure 11C is a graph showing the dynamic COF of micropillars of different densities according to the example embodiment of Figure 11A.

[0037] Figure 12A is a photomicrograph under a 100x optical microscope showing cell samplers with various micropillar patterns prepared according to the example embodiment for optimizing micropillar patterns. The size of the cell sampler was observed under an optical microscope at 100x magnification. The friction of the cell sampler on synthetic skin was measured according to ASTM D1894. The COF of samplers with different patterns was tested and compared.

[0038] Figure 12B is a graph showing the static COF of micropillars with different patterns according to the example embodiment of Figure 12A.

[0039] Figure 12C is a graph showing the dynamic COF of micropillars with different patterns according to the example embodiment of Figure 12A.

[0040] Figure 13A is a photomicrograph under an optical microscope at 100x magnification, showing the cell collection volume assessment of a PDMS sampler after one swab; the photo shows large squares of skin cells / corners: (6+10+2+2) / 4 = 5 collected cells = 5 x 0.1 x 10⁴ = 5000 skin cells (according to the example embodiment). Cells on the sampler were digested with trypsin.Trypan blue staining was performed. Cells were quantified using a hemocytometer.

[0041] Figure 13B is a photomicrograph at 100x magnification showing the cell collection assessment of a PDMS sampler wiped 3 times. The photo shows large squares of skin cells / corners: (10+10+6+6) / 4 = 8 collected cells = 8 x 0.1 x 10⁴ = 8000 skin cells (according to the example embodiment). Other experimental settings are the same as in Figure 13A.

[0042] Figure 13C is a photomicrograph at 100x magnification showing the cell collection assessment of a PDMS sampler wiped 5 times. The photo shows large squares of skin cells / corners: (15+8+10+7) / 4 = 10 collected cells = 10 x 0.1 x 10⁴ = 10000 skin cells (according to the example embodiment). Other experimental settings are the same as in Figure 13A.

[0043] Figure 13D is a photomicrograph under an optical microscope at 100x magnification showing the cell collection assessment of the PDMS sampler after 10 swipes. The photo shows a large square of skin cells / corners: (11+10+9+9) / 4 = 9.75 collected cells = 9.75 x 0.1 x 10⁴ = 9750 skin cells (according to the example embodiment). Other experimental settings are the same as in Figure 13A.

[0044] Figure 13E is a photomicrograph under an optical microscope at 100x magnification showing the cell collection assessment of the PDMS sampler after 15 swipes. The photo shows a large square of skin cells / corners: (15+11+12+14) / 4 = 13 collected cells = 13 x 0.1 x 10⁴ = 13000 skin cells (according to the example embodiment). Other experimental settings are the same as in Figure 13A.

[0045] Figure 13F is a photomicrograph under a 100x optical microscope showing the cell collection amount of the PDMS sampler after 20 swipes, showing large squares of skin cells / corners: (16+14+18+15) / 4 = 15.75 collected cells = 15.75 x 0.1 x 10⁴ = 15750 skin cells (according to the example embodiment). Other experimental settings are the same as in Figure 13A.

[0046] Figure 13G is a graph showing the amount of cells collected relative to the number of swipes in the example embodiment of Figures 13A to 13F, showing that the cell collection amount is proportional to the number of swipes. According to the example embodiment, ultrasonic trypsin treatment further improves cell detachment efficiency compared to trypsin alone.

[0047] Figure 13H is a photograph showing a prototype sampler tool manufactured according to the example embodiment by adding a polypropylene (PP) handle to the head of the PDMS sampler.

[0048] Figure 14A is an amplification diagram of β-globin in Caski cells analyzed by qPCR according to an example embodiment. The Caski cells were collected by a cell sampler and detached by sonicated trypsin treatment. Cell DNA was extracted using Realbest reagent provided by the sponsor.

[0049] Figure 14B is an amplification diagram of β-actin in Caski cells analyzed by qPCR according to an example embodiment. The Caski cells were collected by a cell sampler and detached by sonicated trypsin treatment. Cell DNA was extracted using Realbest reagent.

[0050] Figure 14C is an amplification diagram of GAPDH in Caski cells analyzed by qPCR according to an example embodiment. The Caski cells were collected by a cell sampler and detached by sonicated trypsin treatment. Cell DNA was extracted using Realbest reagent.

[0051] Figure 14D is an amplification diagram of β-globin in human forearm skin cells analyzed by qPCR according to an example embodiment. The human forearm skin cells were collected by a cell sampler and detached by sonicated trypsin treatment. Cell DNA was extracted using Realbest reagent.

[0052] Figure 14E is an amplification diagram of β-actin in human forearm skin cells analyzed by qPCR according to an exemplary embodiment. The human forearm skin cells were collected by a cell sampler and detached by sonication and trypsinization. Cell DNA was extracted using Realbest reagent.

[0053] Figure 14F is an amplification diagram of GAPDH in human forearm skin cells analyzed by qPCR according to an exemplary embodiment. The human forearm skin cells were collected by a cell sampler and detached by sonication and trypsinization. Cell DNA was extracted using Realbest reagent.

[0054] Figure 15A is an amplification diagram of different concentrations of β-actin standard template prepared and analyzed by qPCR according to an exemplary embodiment.

[0055] Figure 15B is a melting curve of different concentrations of β-actin standard template prepared and analyzed by qPCR according to the exemplary embodiment of Figure 15A.

[0056] Figure 15C is a graph showing the Ct values ​​versus DNA copy numbers (logarithmic scale) of different concentrations of β-actin standard template prepared and analyzed by qPCR according to the example embodiment of Figure 15A.

[0057] Figure 15D is a graph showing the melting curves of human forearm skin cell samples prepared and analyzed by qPCR according to the example embodiment.

[0058] Figure 16A is a photograph and micrograph showing CaSki cells used and seeded on a slide, culture plate, and synthetic skin (to simulate cell collection with HPV DNA) according to the example embodiment. Cell samplers were used to wipe the slide and culture plate to collect CaSki cells.Ski cells could not be cultured on synthetic skin, and all cells were suspended in the culture medium instructions, page 4 / 15, 7 CN 122555765 A. HPV-16 was analyzed by qPCR, with β-globin as an internal control.

[0059] Figure 16B is an amplification graph showing HPV-16 and β-globin according to the example embodiment.

[0060] Figure 16C is a melting curve of HPV-16 according to the example embodiment of Figure 16B.

[0061] Figure 16D is a melting curve of β-globin according to the example embodiment of Figure 16B.

[0062] Figure 17A is a photograph of a scaffolded cell sampler (left) and a cell sampler (right) during a sustained 1-month accelerated stability test of the cell sampler at 55°C ± 2°C according to the example embodiment.

[0063] Figure 17B is a graph showing the weight of the cell sampler measured over 1 month according to the example embodiment of Figure 17A.

[0064] Figure 17C is a photomicrograph showing a front view of the scaffolded cell sampler (microcolumn width 140 µm) at 100x magnification, on day 0 (left) and day 30 (right), according to an exemplary embodiment.

[0065] Figure 17D is a photomicrograph showing a front view of the cell sampler (microcolumn width 140 µm) at 100x magnification, on day 0 (left) and day 30 (right), according to an exemplary embodiment.

[0066] Figure 17E is a photomicrograph showing a side view of the microcolumn (microcolumn height 140 µm) of the scaffolded cell sampler at 100x magnification, on day 0 (left) and day 30 (right), according to an exemplary embodiment.

[0067] Figure 17F is a photomicrograph showing a side view of the microcolumn (microcolumn height 140 µm) of the cell sampler at day 0 (left) and day 30 (right) under an optical microscope at 100x magnification, according to an exemplary embodiment.

[0068] Figure 17G is a photomicrograph showing the cell collection volume of the cell sampler evaluated according to an exemplary embodiment. Cells on the sampler were trypsin-digested and stained with trypan blue, and quantified using a hemocytometer under an optical microscope at 100x magnification.

[0069] Figure 17H ​​is a graph showing the cell collection capacity of the sampler as equivalent between month 0 and month 1, according to an exemplary embodiment.

[0070] Figure 18A is a schematic diagram showing the rear dimensions of the exemplary sampler tool according to an exemplary embodiment.

[0071] Figure 18B is a schematic diagram showing the front dimensions of the exemplary sampler tool according to the exemplary embodiment of Figure 18A, where the gray area represents the area containing the exemplary sampler unit. Detailed Implementation Definitions

[0073] As used herein and in the claims, the term "comprising" (or any related form, such as) refers to...The terms “comprise”, “including” (or any related form such as “include”), and “containing” (or any related form such as “contain”) mean including the following elements but excluding others. It should be understood that for each embodiment using the terms “comprising” (or any related form such as “comprise”), “including” (or any related form such as “include”), or “containing” (or any related form such as “contain”), this disclosure / application also includes alternative embodiments in which the terms “comprise”, “including”, or “contain” are replaced with “substantially constitutes” or “consistent with”. These alternative embodiments using “consistent with” or “substantially constitutes” are to be understood as embodiments of a smaller scope than the “comprise”, “including”, or “contain” embodiments.

[0074] For clarity, “comprising,” “including,” “containing,” and “having,” and any related forms, are open-ended terms that allow for additional elements or features beyond the specified essential elements, while “consisting of” is a closed-ended term that limits the elements listed in the claims and excludes any elements, steps, or ingredients not specified in the claims. Specification 5 / 15 pages 8 CN 122555765 A

[0075] As used herein, the singular forms “a / an” and “the” are intended to also include the plural forms unless the context clearly indicates otherwise. When a range is mentioned in the specification, the range is understood to include each discrete point within the range. For example, 1 to 7 means 1, 2, 3, 4, 5, 6, and 7.

[0076] As used herein, the term “about” is understood to be within the normal tolerance range in the art and not exceeding ±10% of the specified value. By way of example only, about 50 means from 45 to 55, including all values ​​in between. As used herein, the phrase “about” includes the specific value, for example, about 50 includes 50.

[0077] It should be understood that terms such as “top,” “bottom,” “middle,” “side,” “length,” “inner,” “outer,” “internal,” “external,” “outer,” “vertical,” and “horizontal” that may be used herein describe reference points only and do not limit the invention to any particular orientation or configuration. Furthermore, terms such as “first,” “second,” and “third” only identify multiple parts disclosed herein.One of the components and / or reference points, and likewise not limiting the present invention to any particular configuration or orientation.

[0078] Although specific embodiments are mentioned in the description, this disclosure should not be construed as limiting it to the embodiments set forth herein. Numbered Embodiments

[0079] Embodiment 1. A cell sampling unit comprising: a plurality of micropillars for contacting a sample, such as skin or other surface, and acquiring cells from the surface of the sample, such as skin or other surface, by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the acquired cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end comprising a contact surface having a frictional force; wherein each micropillar has a diameter of approximately 60-140 µm and a height of approximately 60-140 µm; wherein the plurality of micropillars are configured in an array arrangement; the micropillars are distributed at a density between approximately 1000-1400 micropillars / cm2.

[0080] Example 2. A cell sampling unit as described in Example 1, wherein the height of each micropillar is 60 µm, 100 µm, or 140 µm.

[0081] Example 3. A cell sampling unit as described in any of the preceding examples, wherein the sample, such as skin or other surface, is derived from human skin, such as skin from male genital organs for human papillomavirus (HPV) detection in men.

[0082] Example 4. A cell sampling unit as described in any of the preceding examples, wherein the overall length of the cell sampling unit is approximately 1-2 cm and the width is approximately 1-3 cm.

[0083] Example 5. A cell sampling unit as described in any of the preceding examples, wherein each micropillar is a cylinder, triangular prism, square prism, hexagonal prism, or pentagonal prism.

[0084] Example 6. A cell sampling unit as described in any of the preceding examples, wherein each micropillar and / or the base layer is made of a biocompatible polymer material, such as polydimethylsiloxane (PDMS), thermoplastic polyurethane (TPU), and / or polycaprolactone (PCL).

[0085] Example 7. A cell sampling unit for detecting human papillomavirus (HPV) in men, the cell sampling unit comprising a plurality of micropillars for contacting a sample and acquiring cells on the surface of the sample by friction; and a base layer comprising a front side for supporting the plurality of micropillars and the acquired cells, wherein one end of each micropillar is fixed to or extends from the front side of the base layer, and the other end is a free end comprising a contact surface with friction; wherein each micropillar is cylindrical, with a diameter of approximately 60-140 µm and a height of approximately 100-140 µm.µm; wherein the plurality of micropillars are configured in an array arrangement; the micropillars are distributed at a density of approximately 1000 micropillars / cm2; and wherein each micropillar and / or the base layer is made of polydimethylsiloxane (PDMS).

[0086] Example 8. A cell sampling unit for detection of human papillomavirus (HPV) in men as described in Example 7, wherein the height of each micropillar is 60 µm, 100 µm or 140 µm in the specification 6 / 15 pages 9 CN 122555765 A.

[0087] Example 9. A cell sampling tool comprising: a cell sampling unit as described in any one of Examples 1-8; and an actuator; wherein the base layer of the cell sampling unit is attached to or extends from the actuator, such that the cell sampling unit rubs against the sample under the action of an external force applied to the actuator to obtain cells.

[0088] Example 10. A cell sampling method, the method comprising the steps of: rubbing a sample with a cell sampling unit as described in any one of Examples 1-8 or a cell sampling tool as described in Example 9 to obtain cells therefrom.

[0089] Example 11. The method as described in Example 10, wherein at least 5,000 cells are obtained on the cell sampling unit.

[0090] Example 12. The method as described in Example 10 or 11, wherein more than 25,000 cells are obtained.

[0091] Example 13. The method as described in any one of Examples 10-12, wherein rubbing the sample comprises the steps of: placing the cell sampling unit or the cell sampling tool on a sample surface, such as skin or other surface, the free end facing the sample surface, such as skin or other surface, and rubbing the sample back and forth under the action of an external force to collect the cells.

[0092] Example 14. The method of any one of Examples 10-13, wherein the sample is human skin, such as male genital skin for human papillomavirus (HPV) detection, or skin or surface of other animals or humans.

[0093] Example 15. The method of any one of Examples 10-14, further comprising the steps of: immersing the cell sampling unit or the cell sampling tool in an extraction solution to detach the cells from the sampling unit and causing the cells to detach from the sampler under external force; and separating the detached cells from the extraction solution by centrifugation, removing the supernatant to obtain a cell precipitate suitable for DNA extraction.

[0094] Example 16. The method of any one of Examples 10-15, further comprising the steps of: rubbing the sample, such as human skin, 1-20 times to obtain 5,000-50,000 cells or more. Example Methods, Sampler Head Optimization and Results 1. Fabrication of an example cell sampler head with biomimetic microstructures

[0095] In the examples below, the cell sampler unit may be referred to as a cell sampler head, sampling head, or sampler head. A silicon wafer mold was selected as the master mold for the example cell sampler head. Such a mold has a smooth surface, a high melting point, and dimensions accurate to tens to hundreds of micrometers, making it suitable for manufacturing cell sampler heads using different biocompatible polymers. As shown in Figure 1, the master negative mold is made of a silicon wafer with a microstructure (at least 1400 micropillars / 1.4 cm2). The cell sampler head is cast from PDMS. The microstructure dimensions of the cell sampler head made of PDMS were certified by the supplier using a white light interferometer. The height and width of the cell sampler head manufactured by the supplier are approximately 140 µm. 2. Evaluation of the consistency of the microstructure fabrication of the example cell sampler head using biocompatible polymers.

[0096] In this example, three example biocompatible polymers, including TPU, PDMS, and PCL, were tested as example cell sampler heads. To produce the PDMS cell sampler head, a silicone elastomer kit was used (Figure 2A). 10 parts of silicone base material were mixed with 1 part of curing agent. Air bubbles in the solution were removed by placing it in a desiccator for about 30 min. The clear solution was then cast onto a silicon wafer master mold. The mold containing PDMS was placed in a desiccator to remove air bubbles from the micropillar pores for 18 h, allowing the PDMS to completely fill the micropillar pores. The mold was then placed in an oven at 65°C for 2 h and cut into 1 x 1.4 cm2 pieces. The height and width of the micropillars were measured using an optical microscope. Both the height and width were approximately 140 µm (Figures 2B and 2C).

[0097] For the PCL cell sampler head, a solid PCL was placed on the master mold and incubated in a vacuum oven at 80°C for 18 h (Figure 3A). The sampler head was then cooled to room temperature. The PCL sampler head was carefully removed with a scalpel and cut into approximately 1 x 1.4 cm² pieces, the size of which is shown on page 7 / 15 of the instruction manual, 10 CN 122555765 A. The height and width of the micropillar were measured using an optical microscope. The width of the micropillar was approximately 140 µm (Fig. 3B), and the height was approximately 108 µm (Fig. 3C).

[0098] To manufacture the TPU cell sampler head, TPU particles were placed on a master mold and incubated in a vacuum oven at 200°C for 18 h (Fig. 4A). The sampler head was cooled to room temperature. The TPU sampler head was carefully removed with kitchen scissors and cut into 1 x 1.4 cm² pieces. The height and width of the micropillar were measured using an optical microscope. Both the height and width of the micropillar reached 140 µm (Figs. 4B and 4C).

[0099] The height and width of the micropillar made of PDMS both reached 140 µm. The PDMS cell sampler head is soft and flexible, and is comfortable to wipe the forearm with. It is the best among the three tested materials. Even when incubated in a vacuum oven, the height of the PCL micropillars did not reach 140 mm.µm. This is likely due to its higher viscosity in the liquid state compared to PDMS and TPU. Furthermore, PCL sampler heads are brittle and difficult to wipe on the forearm. It is not considered a material for sampler heads. Although sampler heads made of TPU meet the target size for sampler heads, TPU is difficult to remove from the mold. TPU is somewhat difficult to wipe on the forearm. The production of TPU sampler heads also requires high temperatures. Compared to PDMS, TPU is the second preferred material for sampler heads. The results indicate that PDMS is a relatively better choice for cell sampler head materials due to its texture and ease of production. 3. Characterization of the microstructure of the cell sampler head by scanning electron microscopy (SEM).

[0100] As shown in Figure 5A, an example PDMS cell sampler head was cut and fixed on a metal stage. After gold coating, the microstructure was characterized by SEM, and ten regions of interest (ROI) for each sample were examined. The height and width of the micropillars were 140 µm (Figures 5B and 5C), and details of each measurement are listed in Table 1. The density is also greater than 1000 columns / cm2. Table 1. The microstructure was characterized by scanning electron microscopy and the ten ROIs of each sample were examined. The height and width of all ROIs were approximately 140 µm. Instruction manual 8 / 15 pages 11 CN 122555765 A 4. Optimize the shape, size, density and pattern of the micropillars on the cell sampler head and measure the coefficient of friction on the skin model

[0101] a) Optimize the shape of the micropillars

[0102] In addition to cylindrical columns, square columns and rectangular columns were also prepared. Square and triangular master molds were made (Figs. 6A and 6C). The height and width of the micropillars were 140 µm and designed to be at least 1400 micropillars / 1.4 cm2 (Figs. 6B and 6D).

[0103] The frictional force generated by the microstructure on the synthetic skin was measured by a coefficient of friction (COF) tester (Labthink MXD-02) and expressed as COF. 8N synthetic skin was used as a human forearm and the synthetic skin was fixed to the sliding surface of the tester. The PDMS sampler head was fixed to the center of the sliding sled with double-sided tape, and the weight of the sliding sled with the sampler head was recorded. The test speed was set to 150 mm / min. As shown in Figure 7, the frictional force of the cell sampler head on synthetic skin was measured according to ASTM D1894. The COF of micropillars of different shapes was tested and compared. Typically, the frictional force of the sampler head on human forearm skin is about 0.2-0.5 N. The order of frictional force is as follows: triangular micropillars > circular micropillars > square micropillars > flat surfaces (no micropillars).

[0104] The software "MXD-02 coefficient of friction tester" was used.COF was calculated for cell sampler heads of different shapes. Static friction time was defined as 0-10 seconds, while dynamic COF was calculated based on 10-60 seconds, as shown in Figure 7C. The static and dynamic COF of the sampler heads are shown in Table 2. The percentage increase in COF was calculated as follows: [(COF micropillar - COF flat surface) / COF flat surface * 100%]. Compared to a flat surface without microstructure, the static and dynamic COF of circular micropillars increased by 57.9% and 32.8%, respectively. The static and dynamic COF of triangular micropillars increased by more than 50%, while the static and dynamic COF of square micropillars increased by less than 20%. Based on the COF results, circular and triangular micropillars are relatively better choices for sampler heads.

[0105] For triangular micropillars, different directions of movement may affect COF and cell collection capacity. The COF of triangular micropillars during lateral, forward, and backward movement was tested on synthetic skin using a COF tester (Figure 8A). The frictional force of the cell sampler head on synthetic skin was measured according to ASTM D1894. The COF in different directions of movement ranged from 0.199 to 0.238, with the highest static COF observed when moving backward (Figure 8B). The dynamic COF values ​​ranged from 0.198 to 0.289, with the highest value observed when moving forward (Figure 8C). Table 2. The COF of cell sampler heads of different shapes was measured using a coefficient of friction (COF) tester according to ASTM D1894. The percentage increase in COF was defined as: [(COF micropillar – COF flat surface) / COF flat surface * 100%]. b) Optimization of micropillar size

[0106] To optimize the size of the micropillar, micropillars with heights of 60 µm, 100 µm, and 140 µm were prepared (Figure 9A). Micropillars with a height of 140 µm showed the highest static COF value (Fig. 9B). Micropillars with heights of 100 µm and 140 µm showed the highest dynamic COF (Fig. 9C). In addition, micropillars with widths of 60 µm, 100 µm, and 140 µm were prepared (Fig. 10A). Micropillars with a width of 140 µm showed the highest static COF value (Fig. 10B), while micropillars with a width of 100 µm showed the highest dynamic COF (Fig. 10C). c) Optimization of micropillar density

[0107] Cell sampler heads with densities of 1000, 1200, and 1400 micropillars / cm2 were prepared (Fig. 11A). 1000 micropillars / cm2 showed the highest static and dynamic COF, and the micropillar density was negatively correlated with the dynamic COF (Figs. 11B and 11C). d) Optimization of micropillar pattern

[0108] For patterning, cell sampler heads with micropillar arrays and cross-patterns were prepared (Fig. 12A). The array pattern showed higher static and dynamic COF than the cross-pattern (Figs. 12B and 12C).

[0109] In summary, cell samplers 1, 3, and 4 all exhibited high dynamic COF values ​​on synthetic skin (Table 3). Specification 10 / 15 pages 13 CN 122555765 A Table 3. Dynamic coefficient of friction for samplers of different sizes. n = 3. 5. Evaluation of the ability of the cell sampler prototype to collect skin cells from the human forearm.

[0110] The sampler tool prototype was manufactured by adding a polypropylene (PP) handle to the sampler head (Fig. 13H). Briefly, the master silicone mold was placed in a soft silicone mold. The PP handle was inserted into the soft mold, and PDMS was poured into the mold. The mold was degassed in a desiccator and incubated at 65°C as described in Section 2. The sampler head was then cut to a size of 1 x 1.4 cm2.

[0111] The forearm was wiped 1, 3, 5, 10, 15, and 20 times. The sampler head was placed in a 50 ml tube, and 2 ml of trypsin was added to completely submerge the sampler head, followed by incubation at 37°C for 3 min. The sampler head was then removed from the solution, and the solution was centrifuged at 3000 rcf for 5 min. The supernatant was carefully removed using a pipette. 50 µl of Tris-buffered saline (TBS) was added to the bottom of the tube, followed by 50 µl of trypan blue. 10 µl of the solution was transferred to a hemocytometer, and the cell count was performed under an optical microscope. The average number of cells on the four large squares at the corners was measured (Fig. 13A-F). Our data indicate that approximately 5000–15000 cells were collected after different numbers of wipings. The number of cells collected was proportional to the number of wipings (Fig. 13G).

[0112] To further improve cell detachment, sonication with trypsin was performed. After collecting skin cells, the sampler tip was placed in a 15 ml tube, and 2.5 ml of trypsin was added to completely submerge the tip. The tube was then incubated and sonicated at 37°C in a temperature-controlled sonicator for 10 min. The sampler tip was then removed from the solution, and approximately 1.4 ml of sample solution was transferred to a 1.5 ml tube. The sample solution was centrifuged at 20,000 rcf for 5 min. The supernatant was carefully removed using a pipette, and the remaining solution (approximately 1.1 ml) in the 15 ml tube was transferred to the 1.5 ml tube. The sample solution was centrifuged again at 20,000 rcf for 5 min. After carefully removing the supernatant, 50 µl of TBS was added to the precipitate. Then, 50 µl of trypan blue was added to the 1.5 ml tube. 10 µl of the solution was transferred to a hemocytometer, and the cell count was performed under an optical microscope. Approximately 25,000–48,000 cells were collected through various wiping cycles. Therefore, sonication with trypsin was used as a cell detachment method.

[0113] The cell collection capabilities of sampler heads 1, 3, and 4 were examined. The cell collection volumes of sampler heads 1, 3, and 4 were comparable. Sampler head 1 was in use, therefore sampler head 1 was selected for further development. 6. Evaluate the amount of genomic DNA extracted from skin cells collected by the prototype cell sampler. Instruction manual 11 / 15 pages 14 CN 122555765 A

[0114] Genomic DNA was extracted from cells collected by the cell sampler using a commercially available kit (Qiagen). Hold the sampler and swab it on your forearm 20 times. Place the sampler head in a 50 ml tube and add 2 ml of trypsin to the sampler head. Incubate the tube at 37°C for 3 min. Then remove the sampler head from the solution and centrifuge the solution at 3000 rcf for 5 min. Carefully remove the supernatant using a pipette. Add 200 µl PBS, 20 µl proteinase K, and 200 µl buffer AL to the tube. Mix the solution thoroughly and incubate at 56°C for 10 min with oscillation at 300 rpm. Cool the solution to room temperature and add 200 µl ethanol. Transfer the solution to a DNeasy Mini spin column and place it in a 2 ml collection tube. Centrifuge the sample at 6000 rcf for 1 min. Discard the flow-through in the collection tube. Then place the spin column in a new 2 ml collection tube. Add 500 µl buffer AW1 to the spin column and centrifuge at 6000 rcf for 1 min. Discard the flow-through in the collection tube. After placing the spin column in a new 2 ml collection tube, add 500 µl buffer AW2 to the spin column. Centrifuge the column at 20000 rcf for 3 min. Discard the flow-through and transfer the spin column to a new 1.5 ml tube. Elute the DNA by adding 50 µl buffer AE to the center of the spin column membrane. The column was incubated at room temperature (15°C–25°C) for 1 min. The column was centrifuged at 6000 rcf for 1 min.

[0115] DNA quantification was performed using NanoDrop (Implen NP80) by UV absorbance at 260 nm. Briefly, 1 µl of buffer AE was pipetted into NanoDrop and calibrated as a blank. Then 1 µl of filtrate was pipetted into NanoDrop for DNA measurement at a concentration of ng / ml. As shown in Table 4, when cells were detached from the cell sampler head using trypsin, the amount of DNA extracted per cell sampler head was greater than 4 ng / µl, while when cells were detached using trypsin with sonication, the amount of DNA extracted per cell sampler head increased to 5.67 ng / µl. Table 4. Sampler head swabbing human forearm skin and detaching cells with trypsin or trypsin with sonicationCells. Genomic DNA was extracted from cells collected by a cell sampler head using a commercially available kit (Kiager). The DNA concentration of the extraction filtrate was quantified using UV absorbance at 260 nm. 7. Evaluation of the copy number of housekeeping genes in the extracted DNA by real-time quantitative PCR

[0116] After genomic DNA was extracted using Realbest reagent (provided by the sponsor), the copies of housekeeping genes present in the sample were obtained by real-time quantitative PCR (qPCR). Briefly, the collected cells were detached by trypsin and sonication. The collected cell pellet was then resuspended in 50 µl of Realbest reagent and incubated at 98°C for 30 min with shaking at 300 rpm. The sample solution was centrifuged at 8000 rpm for 5 min. 40 µl of the supernatant was transferred to a new tube. The extracted DNA was mixed with the qPCR master mixture and the primers for the housekeeping genes. The sample was then processed in a qPCR thermal cycler. Plasmids containing housekeeping genes were used to establish a standard curve. The copy number of housekeeping genes in the extracted DNA samples was calculated based on the generated standard curve.

[0117] Initially, β-globin was proposed as a housekeeping gene. However, β-globin was detected in CaSki cells, but not in forearm skin cells. Other common housekeeping genes, β-actin and GAPDH, were also tested. Both β-actin and GAPDH were detected in both CaSki cells and forearm skin cells (Figures 14A-14F). β-actin had a higher CT value than GAPDH and was therefore selected as the housekeeping gene for internal control in human skin cells.

[0118] The copy number of housekeeping genes in cells collected from human forearm skin was calculated. A β-actin standard template was used to establish a standard curve (Figures 15A and 15C). The melting curve confirmed the specificity of the β-actin standard (Figure 15B).

[0119] After wiping the human forearm 20 times with a sampler, the gene copy number of the housekeeping gene in the extracted DNA samples was calculated (Table 5). The CT values ​​of all samples were approximately 30-31, and the average gene copy number was approximately 2200 / µl. Table 5. Gene copy number of human β-actin collected from human forearm and analyzed by qPCR. 8. Simulation of cell collection with HPV DNA using cell lines with HPV DNA

[0120] Since it was not possible to obtain HPV-infected cells from healthy volunteers, an alternative method was used. The previously HPV-infected human cancer cell line, Ca Ski cells, was used. Cells were seeded at a density of 5 x 10⁴ cells / mm² on slides, culture plates, and synthetic skin, a density similar to the epidermal cell density of the human forearm (Figure 16A). Ca Ski cells were notCells were cultured on synthetic skin and suspended in culture medium. The sampler was used to wipe slides and culture plates 20 times. Cells were detached by sonication and trypsin treatment. DNA was extracted using a Qiager DNA Extraction Kit. HPV-16 was analyzed by qPCR, with β-globin as an internal control. Individual CaSki cells served as a positive control. On the amplification plot, the CT values ​​for both HPV-16 and β-globin were below 40, indicating the presence of HPV DNA (Figure 16B). Melting curves also confirmed the specificity targeting HPV DNA (Figure 16C). Instructions for Use, Pages 13 / 15, CN 122555765 A, Table 6. HPV-16 positive cell line CaSki cells were seeded on slides and culture plates to simulate cell collection with HPV DNA. HPV-16 was analyzed by qPCR, with β-globin as an internal control. 9. Accelerated stability testing of the cell sampler was performed at 55°C ± 2°C for 1 month to assess its cell collection capacity.

[0121] The stability of the cell sampler prototypes was evaluated under accelerated aging conditions. The aging factor was 2³ = 8 under accelerated aging conditions at 55°C above ambient temperature (above 25°C). The cell sampler prototypes (with and without scaffolds, Fig. 17A) were stored for one month (31 days) in a temperature-controlled chamber at 55°C ± 2°C and atmospheric humidity, and cell collection capacity was evaluated as described in Section 5 (Ultrasonic Trypsin Treatment). The weight of the cell samplers was measured for one month, and the weight of all samplers remained stable throughout the month (Fig. 17B). The dimensions of the micropillars on the samplers were monitored. No changes in the width and height of the samplers were observed after one month in all samplers (Figs. 17C, 17D, 17E, and 17F). The width and height of all samplers (Figs. 17C and 17D) remained at approximately 140 µm. The cell collection volume of the cell samples was evaluated. Cells on the samplers were trypsin-digested and stained with trypan blue. Cells were quantified using a hemocytometer under an optical microscope. According to Figure 17H, no significant difference in the cell collection capacity of the sampler was observed between month 0 and month 1. 10. Example Sampling Tool

[0122] Figure 13C shows an example sampling tool comprising a handle and a cell sampling unit. The sampler tool prototype is manufactured by attaching a polypropylene (PP) handle to the PDMS sampler head.

[0123] Figures 18A and 18B show another example sampler tool containing any of the example cell sampler units as described herein. In this example, the example cell sampler unit is formed as a single piece with the manipulator. The gray area represents the region containing the example sampler unit, located at the front end of the manipulator. The manipulator has two curved sections to facilitate the user's grip to apply external force to the manipulator, thereby applying force to the sampler unit. Conclusion

[0124] In summary, examples of the provided biocompatible polymer-based cell sampler unit and cell sampler have been developed, which can be used to efficiently and painlessly collect cells for male HPV testing. In some embodiments, the provided cell sampler is small and flexible. In some embodiments, the size of the cell sampler head is approximately 1 x 1.4 cm². In some embodiments, the sampling surface contains a regular array of >1000 micropillars / cm², each micropillar having a height and width of <150 µm. Due to the novel design, the cell sampler is able to collect a sufficient number of cells for downstream HPV analysis. After collecting the cells (see page 14 / 15 of the instruction manual, CN 122555765 A), the cell sampler is enzymatically digested to detach the cells. The detached cells are used for DNA extraction and further qPCR analysis.

[0125] The provided example cell sampler is an easy-to-use, convenient, and reliable sampling device capable of self-sampling or remote sampling. Given a specified size and microstructure, the sampler can be used not only for HPV testing but also for other diagnostic tests, such as those for dermatology. Instruction Manual Page 15 / 15 18 CN 122555765 A Figure 1A Figure 1B Instruction Manual Drawing Page 1 / 35 19 CN 122555765 A Figure 1C Figure 1D Instruction Manual Drawing Page 2 / 35 20 CN 122555765 A Figure 2A Instruction Manual Drawing Page 3 / 35 21 CN 122555765 A Figure 2B Figure 2C Instruction Manual Drawing Page 4 / 35 22 CN 122555765 A Figure 3A Instruction Manual Drawing Page 5 / 35 23 CN 122555765 A Figure 3B Figure 3C Instruction Manual Drawing Page 6 / 35 24 CN 122555765 A Figure 4A Instruction Manual Drawing Page 7 / 35 25 CN 122555765 A Figure 4B Figure 4C Instruction Manual Drawing Page 8 / 35 26 CN 122555765 A Figure 5A Instruction manual figures 9 / 35, page 27, CN 122555765 A, Figure 5B, Figure 5C, Figure 6A; Instruction manual figures 10 / 35, page 28, CN 122555765 A, Figure 6B, Figure 6C, Figure 6D; Instruction manual figures 11 / 35, page 29, CN 122555765 A, Figure 7; Instruction manual figures 12 / 35, page 30, CN 122555765 A, Figure 8A, Figure 8B; Instruction manual figures 13 / 35, page 31, CN 122555765 A, Figure 8C, Figure 9A; Instruction manual figures.Page 14 / 35, 32 CN 122555765 A, Figure 9B, Figure 9C, Instruction Manual Drawings; Page 15 / 35, 33 CN 122555765 A, Figure 10A, Figure 10B, Instruction Manual Drawings; Page 16 / 35, 34 CN 122555765 A, Figure 10C, Figure 11A, Instruction Manual Drawings; Page 17 / 35, 35 CN 122555765 A, Figure 11B, Figure 11C, Instruction Manual Drawings; Page 18 / 35, 36 CN 122555765 A, Figure 12A, Figure 12B, Instruction Manual Drawings; Page 19 / 35, 37 CN 122555765 A, Figure 12C, Figure 13A, Instruction Manual Drawings; Page 20 / 35, 38 CN 122555765 A, Figure 13B, Figure 13C, Figure 13D, Instruction Manual Drawings; Page 21 / 35, 39 CN 122555765 A, Figure 13E. Figure 13F: Instruction manual drawing, page 22 / 35, 40 CN 122555765 A Figure 13G: Instruction manual drawing, page 23 / 35, 41 CN 122555765 A Figure 13H: Instruction manual drawing, page 24 / 35, 42 CN 122555765 A Figure 14B: Instruction manual drawing, page 25 / 35, 43 CN 122555765 A Figure 14E: Instruction manual drawing, page 14F: Instruction manual drawing, page 26 / 35, 44 CN 122555765 A Figure 15B: Instruction manual drawing, page 27 / 35, 45 CN 122555765 A Figure 15D: Instruction manual drawing, page 28 / 35, 46 CN 122555765 A Figure 16A: Instruction manual drawing, page 29 / 35, 47 CN 122555765 A Figure 16B: Figure 16C Instruction Manual Drawings, Page 30 / 35, 48 CN 122555765 A Figure 16D Figure 17A Instruction Manual Drawings, Page 31 / 35, 49 CN 122555765 A Figure 17B Figure 17C Figure 17D Instruction Manual Drawings, Page 32 / 35, 50 CN 122555765 A Figure 17E Figure 17F Figure 17G Instruction Manual Drawings, Page 33 / 35, 51 CN 122555765 A Figure 17H ​​Figure 18A Instruction Manual Drawings, Page 34 / 35, 52 CN 122555765 A Figure 18B Instruction Manual Drawings, Page 35 / 35, 53 CN 122555765 A

Claims

1. A cell sampling unit comprising: Multiple micropillars, the multiple micropillars being used to contact a sample and acquire cells from the surface of the sample through friction; and The base layer includes a front side for supporting the plurality of micropillars and the acquired cells. Each micropillar has one end fixed to or extending from the front side of the base layer, and the other end is a free end containing a contact surface with friction. Each micropillar has a diameter of approximately 60-140 µm and a height of approximately 60-140 µm. The plurality of micropillars are configured in an array arrangement; the micropillars are arranged at a density of approximately 1000-1400 micropillars / cm. 2 Density distribution between them.

2. The cell sampling unit of claim 1, wherein the sample is derived from human skin, such as skin from male reproductive organs for the detection of human papillomavirus (HPV) in men.

3. The cell sampling unit as claimed in claim 1, wherein the overall length of the cell sampling unit is approximately 1-2 cm and the width is approximately 1-3 cm.

4. The cell sampling unit as described in claim 1, wherein each micropillar is a cylinder, triangular prism, square prism, hexagonal prism, or pentagonal prism.

5. The cell sampling unit of claim 1, wherein each micropillar and / or the substrate is made of a biocompatible polymer material, such as polydimethylsiloxane (PDMS), thermoplastic polyurethane (TPU), and / or polycaprolactone (PCL).

6. A cell sampling unit for detecting human papillomavirus (HPV) in men, the cell sampling unit comprising... Multiple micropillars are used to contact the sample and acquire cells on the surface of the sample through friction; as well as The base layer includes a front side for supporting the plurality of micropillars and the acquired cells. Each micropillar has one end fixed to or extending from the front side of the base layer, and the other end is a free end containing a contact surface with friction. Each micropillar is cylindrical, with a diameter of approximately 60-140 µm and a height of approximately 100-140 µm. The plurality of micropillars are configured in an array arrangement; the micropillars are arranged at approximately 1000 micropillars / cm. 2 The density distribution; and Each micropillar and / or the substrate is made of polydimethylsiloxane (PDMS).

7. A cell sampling tool, comprising: The cell sampling unit as described in any one of claims 1-6; and Operator; The base layer of the cell sampling unit is attached to or extends from the manipulator, such that the cell sampling unit rubs against the sample to obtain cells under the action of an external force applied to the manipulator.

8. A cell sampling method, the method comprising the following steps: Cells are obtained by rubbing a sample with the cell sampling unit as described in any one of claims 1-6 or the cell sampling tool as described in claim 7.

9. The method of claim 8, wherein the friction sample comprises the following steps: The cell sampling unit or the cell sampling tool is placed on the sample surface, with the free end facing the sample surface, and the sample is rubbed back and forth under the action of an external force to collect the cells.

10. The method of claim 8 or claim 9, wherein the sample is human skin, such as male genital skin used for human papillomavirus (HPV) detection in men.

11. The method of any one of claims 8-10, further comprising the following steps: The cell sampling unit or the cell sampling tool is immersed in the extraction solution to detach the cells from the sampling unit and to induce the cells to detach from the sampler under external force; and The detached cells are separated from the extract by centrifugation, and the supernatant is removed to obtain a precipitate that can be used for DNA extraction.

12. The method of any one of claims 8-10, further comprising the following steps: Rub the sample, such as human skin, 1-20 times to obtain 5,000-50,000 cells or more.

Images