Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Overview of microfluidic-based tumor models using patient-derived materials.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Chemotaxis Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Microfluidic systems using patient-derived materials for chemotherapy (A–G) and immunotherapy (H,I) . (A) Top: schematic of a patient-derived tumor explant, microdissected, loaded in a microfluidic device and injected with various drugs. Results are then interpreted to identify non-responders to treatment for a personalized treatment strategy. Bottom: photograph of tumor prior and after microdissection in 500 μm-PDMEs and design of the microfluidic device containing up to 5 chambers for PDME entrapment and drug testing. (B) PDMEs from various patients and different cancers are analyzed by live/dead cell assay using confocal microscopy (green = live, red = dead), showing various PDME architectures and viability patterns. (C) PDMEs from an ovarian tumor treated with carboplatin shows high heterogeneity in PDME structure and viability response for PDME originating from the same tumor. (D) Corresponding graphs of viability data from (B) (top) and (C) (bottom) compared to non-treated controls show inter-cancer tumor heterogeneity and drug efficacy on PDMEs from one tumor, despite intra-tumor heterogeneity observed in (C) . (E) Microfluidic device enabling in situ spheroid patterning technique: a microfluidic chamber (i) is filled with a mixture (blue) containing hydrogel precursors, photoinitiator, and patient-derived tumor cells (ii) and then illuminated with UV light through a photomask (gray) (iii). Exposed precursor is crosslinked into a hydrogel (dark blue), detaining cells within the region (iv), and non-crosslinked gel is flushed form the chamber with clean saline from the chamber (v). Finally, saline is replaced with media (red) (vi) for incubation. (F) Schematic of system that offers low volumes and closed loop fluidic circuit controlled by a computer-controlled peristatic pump to treated individual organoids. (G) In vitro chemotherapy assessment of organoids derived from two patients with mesothelioma treated with various drug doses and combinations shows different response between patients, although treatment occurred at different times between patients (viability measured from live/dead confocal microscopy on organoids). (H) Schematic of PDMEs obtained after dissection and sieving. Fraction 2 (40–100 μm) is usually used in microfluidic devices such as the 3D cell culture chip from AIM Biotech shown, which contains a center gel region. Gel loading port and media ports labeled (B) , along with center and side channels (C) . (I) The AIM Biotech microfluidic device was used by Wang et al. to test a selective RIP1 inhibitor on pancreatic ductal adenocarcinoma PDMEs. After RIP1i treatment ( n = 5 patients shown), PDME live/dead ratio, and size decreased. (A–I) reproduced with permission from Astolfi et al. , Aref et al. , Mazzocchi et al. , and Wang et al. , respectively. (D) Top: **Result for PDMEs stained and imaged a second time; Bottom: * p -value = 0.014. (G) * p < 0.05, ** p < 0.01. (I) * p < 0.05.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Derivative Assay, Injection, Laser Capture Microdissection, Confocal Microscopy, In Situ, Incubation, In Vitro, Dissection, Cell Culture, Labeling, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Addressing Patient Specificity in the Engineering of Tumor Models

doi: 10.3389/fbioe.2019.00217

Figure Lengend Snippet: Tumor-on-chip systems. (A) Diagram illustrating the processing steps involved in the preparation of patient-derived tumor cells and tumor-infiltrating lymphocytes (TILs) from the same tumor sample, and live imaging by confocal microscope. (B) 12-channel multiplexed cyclic-olefin-copolymer (COC)-based microfluidic device with V-trap design for capturing tumor sample in flow stream and dual port entry for TILs. (C) A 3D multicellular tumor microenvironment microfluidic model consisting of a middle hydrogel channel (2) surrounded by two media channels (1, 3) for the mechanistic study of the effect of monocytes on T cell receptor-redirected T cell (TCR T cell) killing of tumor cell aggregates. Human monocytes were inserted together with target HepG2-preS1-GFP cell organoids in collagen gel in the central hydrogel region (2), while hepatitis B virus (HBV)-specific TCR T cells were added into one fluidic channel (1) to mimic the intrahepatic carcinoma environment. (D) Representative confocal image of a target cell organoid (in green) surrounded by monocytes (in blue) and HBV-specific TCR T cells (in white), in which the presence of dead target cells is DRAQ7+. (E) Left: Representative target cell HepG2 cell organoids (Hep) cultured with monocytes (Mo) and/or HBV-specific T cell (Ts), in which the presence of dead target cells is DRAQ7+(in red). HBV-specific TCR T cells are labeled with Cell tracker violet dye (in white), while monocytes are unlabeled. Right: Box plot of the percentage of dead target volume after 24 h of co-culture with retrovirally transduced (Tdx) HBV-specific TCR T cells (Tc = control T cell). (F) Metastasis-on-a-chip device and in situ tumor and tissue construct biofabrication. Arrows show fluid flow (E, endothelium; Lu, Lung; C, colorectal cancer organoids made of RFP tagged HCT116 cells; Li, liver; blank, control). Constructs comprised of cells in ECM hydrogels exist under fluid flow and have the capability to experience circulating cells either interact or pass by. (G) Metastasis tracking at day 1 and day 15 showing HCT116 cells colonizing other organs, using phase and epifluorescence microscopy (A–G) reproduced with permission from Lee et al. , Moore et al. , and Aleman and Skardal , respectively. * P ≤ 0.05, *** P ≤ 0.001.

Article Snippet: The spheroid fraction was mixed with collagen-I and used for culture in a cyclic olefin co-polymer (COC)-based 3D microfluidic device (DAX-1, AIM BIOTECH) (Aref et al., ).

Techniques: Derivative Assay, Imaging, Microscopy, Cell Culture, Labeling, Co-Culture Assay, In Situ, Construct, Epifluorescence Microscopy

Journal: Scientific Reports

Article Title: Radiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture

doi: 10.1038/s41598-022-11023-z

Figure Lengend Snippet: The graphical abstract of the study. Patients were classified into two groups: the Control group (only surgery) and the Test group (surgery + IORT). 3D experiments: On day 0, ( A , B ) Mechanically and enzymatically dissociation of the tumor specimens, respectively. ( C , D ) Filtration of the dissociated specimens using 100 µm and 40 µm cell strainers, respectively. ( E ) Embedding the spheroids into prepared collagen gel solution. ( F ) Filling the gel (collagen + spheroids) into the central channels and RPMI + FBS into media channels of the microfluidic devices. On days 1–6, I: Control device whose side channels are loading up with RPMI + FBS. II: Test device whose media channels are loading up with 24 h-wound fluid (WF/WF-RT). Optical imaging and media replacement were accomplished from day 0 to day 6 (RPMI + FBS for control devices and 24 h-wound fluid for test devices). On day 6, Live/Dead staining, immunocytochemistry, and fluorescent imaging. 2D experiments: assays on breast cancer cell lines under 24 h-WF/WF-RT treatment. The figure was created using Biorender ( https://biorender.com ).

Article Snippet: Microfluidic chips were designed and manufactured at AIM BIOTECH (DAX-1, AIM BIOTECH, https://www.aimbiotech.com ) – .

Techniques: Control, Filtration, Optical Imaging, Staining, Immunocytochemistry, Imaging

Journal: Scientific Reports

Article Title: Radiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture

doi: 10.1038/s41598-022-11023-z

Figure Lengend Snippet: Optical and fluorescent images of spheroids in microfluidic devices on days 0, 1, 3, and 6 of culture. ( A ) Tumor spheroids are observed with a convert microscope before mixing with collagen at day 0. Scale bar: 100 µm. Original magnification: ×40. ( B ) Tumor spheroids stained with AO/PI before injection into the microfluidic devices to find spheroid viability on day 0. Scale bar: 100 µm. Original magnification: ×40. ( C ) The tumor-derived spheroids were treated with WF-RT on days 0, 3, and 6 with inverted phase-contrast microscopy and fluorescent microscopy. Different behaviors of three types of spheroids treated with WF-RT were traced for 6 days and shown with the circular dotted line. White dotted lines show motility of the spheroids during 6 days, while green ones represent the in-situ proliferation of cells within the spheroids and pink ones represent the spheroid without proliferation and motility. Scale bars: 100 µm. Original magnification: ×40. ( D ) live/dead staining of the spheroids under RPMI treatment comparing wound fluid treatment in a control sample (spheroids from the non-IORT treated patient) and a test sample (spheroids from IORT treated patient). Scale bars: 100 µm. Original magnification: ×40. Green: AO/live cells; Red: PI/dead cells. ( E ) The graph presents the comparison of %live and %dead cells in control samples with test samples and between RPMI and WF treated cells in each group. Control group: patients who only went under surgery, test group: patients who received IORT during the surgery. CTR control (RPMI + 10%FBS), WF wound fluid, WF-RT IORT-treated wound fluid. ns: non-significant. ***P < 0.001.

Article Snippet: Microfluidic chips were designed and manufactured at AIM BIOTECH (DAX-1, AIM BIOTECH, https://www.aimbiotech.com ) – .

Techniques: Microscopy, Staining, Injection, Derivative Assay, In Situ, Control, Comparison

Journal: Scientific Reports

Article Title: Radiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture

doi: 10.1038/s41598-022-11023-z

Figure Lengend Snippet: Effects of WFs on migration and invasion BC cell line (MDA-MB 231) and in human-derived tumor spheroids. ( A ) Images and graphs related to scratch assay (wound healing compared to negative control in 0 and 24 h). The graph presents the percentage of migrated cells. ( B ) Images of the cells on the upper chamber in transwell assay and graph present the percentage of cell migration. *P < 0.05, and **P < 0.01. ( C ) Migration of BC tumor spheroids in microfluidic devices. Circular dotted lines show one of the spheroids that migrate during 6 days of incubation with WF. Scale bars: 100 µm. Original magnifications: ×40.

Article Snippet: Microfluidic chips were designed and manufactured at AIM BIOTECH (DAX-1, AIM BIOTECH, https://www.aimbiotech.com ) – .

Techniques: Migration, Derivative Assay, Wound Healing Assay, Negative Control, Transwell Assay, Incubation