Table 1. Summary of Spheroid Fabrication Techniques Used in Microfluidic Systemsa.
| technique | cell type used | biomaterial type used | obtained spheroid size/volume/area | no. of fabricated spheroids/spheroid formation time | application area | reference |
|---|---|---|---|---|---|---|
| droplet | rat hepatoma continuous cell line (H4-II-EC3) | agarose | 72.9 ± 18.6 μm | 500 spheroids in 11 h | 3D cell culture | (61) |
| murine colorectal carcinoma cell line (CT26.WT) | alginate | up to 0.1 mm3 | >1000 droplets/s | anticancer therapies | (62) | |
| human cervical carcinoma cells (HeLa) | Matrigel and alginate | 138 ± 20 μm | NR/96 h | anticancer therapies | (63) | |
| human mesenchymal stem cells (hMSC) | alginate/arginine-glycine-aspartic acid (-RGD) | 30–80 μm | NR/150 min | tissue engineering | (64) | |
| diffuse large B-cell lymphoma cell line (SUDHL-10); fibroblast cell line (HS-5); peripheral blood mononuclear cells (PBMCs) | alginate/PuraMatrix | 350 ± 25 μm | 250 spheroids/NR | drug screening | (65) | |
| embryo-derived teratocarcinoma cell line (P19) | alginate | 111 μm | NR/48 h | tissue engineering | (66) | |
| human breast adenocarcinoma cell line (MCF-7); human mammary fibroblast cells (HMF) | alginate | NR | 200 spheroids/min /7 days | drug screening | (67) | |
| primary human bone-marrow-derived mesenchymal stem cells (hBMSC) | polyethylene glycol-diacrylic (PEGDA) | <50 μm | NR/28 days | 3D cell culture | (68) | |
| human glioblastoma cells (U87MG)/mouse neural stem cells (NE-4C) | 100–130 μm | 42,000 spheroids/1 h | regenerative therapy | (69) | ||
| human glioblastoma cell line (U-251) | polyethylene glycol (PEG)/RGDs | 118–480 μm | NR/1 h | drug screening | (70) | |
| hMSC | poly(vinyl alcohol) (PVA) | 90 μm | NR/4 weeks | drug screening | (71) | |
| MCF-7/human fibroblast cell line (HS-5) | alginate | 170 μm | 1000 spheroids/48–72 h | anticancer therapies | (72) | |
| human embryonic kidney cells (HEK293)/human bladder cancer cell line (RT4)/human epidermoid carcinoma cells (A431) | PEG–perfluoropolyether (PEG–PFPE) | NR | 85,000 spheroids/1 h | 3D cell culture | (73) | |
| electrowetting | Madin-Darby canine kidney epithelial cells (MDCK) | Geltrex/agarose/polyacrylamide/alginate/type I collagen | 20 μm | NR/1 day | 3D cell culture | (74) |
| HepG2/mouse embryonic fibroblast cells (NIH-3T3) | type I collagen | NR | NR | drug screening | (75) | |
| mouse bone-marrow-derived mesenchymal stem cells (BM-mMSC)/ HT-29 | up to 400 μm | NR/in 72 h | 3D cell culture | (76) | ||
| microwell | HeLa/human umbilical vein endothelial cells (HUVECs) | PEG | 50–300 μm | NR/up to 36 h | anticancer therapies | (77) |
| human dermal fibroblasts (hDFs) | cellulose nanocrystals and gelatin | ∼150 μm | 2400 spheroids/5 day | drug screening | (78) | |
| MCF-7 | agarose | 200–600 μm | up to 175 spheroids/72 h | drug screening | (79) | |
| human lung carcinoma epithelial cell line (A549)/ human osteoblasts/patient-derived spine metastases cells (BML) | up to 250 μm | 10 spheroids/5 day | personalized medicine | (80) | ||
| human high-grade glioma cells (UVW)/human prostate cancer cell line (LNCaP)/patient biopsy-derived prostate cancer cells | 50–150 μm | 240 spheroids/48 h | personalized medicine | (81) | ||
| rat embryonic fibroblast cells (REF52)/Madin-Darby canine kidney (MDCK) cells | fibronectin/collagen | 40–100 μm | NR | tissue engineering | (82) | |
| human colorectal adenocarcinoma cell line (HT29) | up to 250 μm | 25 spheroids/7 days | drug screening | (83) | ||
| human colorectal carcinoma cell line (HCT116)/human glioma cell line (U87) | 150–200 μm | 50 spheroids/48 h | drug screening | (84) | ||
| HT-29 cells | 130–250 μm | 20 spheroids/6 days | anticancer therapies | (85) | ||
| MCF-7/U87 | agarose | up to 500 μm | 40 spheroids/5 days | anticancer therapies | (86) | |
| human hepatocellular carcinoma cells (HepG2-C3A) | gelatin methacryloyl (GelMA) | 191 ± 10 μm | 10,000 spheroids/5 days | drug screening | (87) | |
| human lung adenocarcinoma cells (A549)/human lung fibroblast cells (MRC-5) | type I collagen | NR | 28 spheroids/72 h | regenerative therapy | (88) | |
| human articular cartilage cells (hACs) | NR | NR/14 days | tissue engineering | (13) | ||
| murine ES cell (ES-D3)/human hepatocellular carcinoma cell (HepG2)/monkey kidney epithelial fibroblast (COS-7) | up to 210 μm | 5000 spheroids/24 h | 3D cell culture | (89) | ||
| human glioma cell line (U87) | PEGDA | 361.3 ± 36.2 μm | 24 spheroids/24–48 h | drug screening | (90) | |
| human mesenchymal stem cells (hMSC) | chitosan/polydopamine | up to 500 μm | NR/5 days | 3D cell culture | (91) | |
| human metastatic breast adenocarcinoma cell line (MDA-MB-231)/human nontumorigenic mammary epithelial cell line (MCF-10A) | type I collagen | 100 μm | 1296 spheroids/5–6 days | 3D cell culture | (92) | |
| MCF-7/HCT-116 | 180 μm | 240 spheroids/24 h | 3D cell culture | (93) | ||
| human colon adenocarcinoma cell line (Caco-2)/normal human dermal fibroblasts (NHDF)/human alveolar basal adenocarcinoma cell line (A549)/human hepatocellular carcinoma cell line (HepG2) | up to 828.7 ± 49.5 μm | 360 spheroids/72 h | tissue engineering and drug screening | (94) | ||
| human hepatoma cells (Huh-7) | Geltrex | 160 μm | 120 spheroids/24 h | drug screening | (95) | |
| A549/human fetal lung fibroblast cell line (MRC-5) | type I collagen | 142.3 μm2 | 84 spheroids/24 h | tissue engineering | (96) | |
| hanging drop | mouse embryonic stem cells (mESC)/human lung cancer cell line (A541)/human leukemia cell line (HL-CZ) | up to 250 μm | 234 spheroids/24 h | 3D cell culture | (97) | |
| mouse embryonic stem cells (mESCs) and human MDA-MB-231 and MCF-7 breast cancer cells | Matrigel | up to 730 ± 27 μm | 16 spheroids/10 days | 3D cell culture | (98) | |
| human epithelial carcinoma cell (A431.H9) | up to 0.16 μL | 384 spheroids/7 days | drug screening | (99) | ||
| mouse embryonic stem cells (mESCs; ES-D3) | 80–120 μm | 72 spheroids/24 h | tissue engineering | (100) | ||
| human Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC) | up to 500 μm | 24 spheroids/7 h | 3D cell culture | (101) | ||
| human umbilical cord blood-derived MSCs/mouse podocyte cells | up to 500 μm | 49 spheroids/24 h | tissue engineering | (102) | ||
| human glioblastoma cell lines (LN229 and PDX) | 4 nL | 900 spheroids/1 h | drug screening | (103) | ||
| human synovial sarcoma-derived cell line (HS-SY-II)/human umbilical cord-derived mesenchymal stem cells (UC-MSCs) | NR | NR/24 h | anticancer therapies | (104) | ||
| human hepatocellular carcinoma cell line (MHCC97-H) | 522 ± 40 μm | 26 spheroids/3 days | 3D cell culture | (105) | ||
| microstructures | HepG2/mouse fibroblast cells (Balb/c-3T3) | PEGDA | NR | 56 spheroids/48 h | drug screening | (106) |
| MCF-7/HepG2 | up to 1 × 10–2 mm3 | 16 spheroids/2 days | drug screening | (107) | ||
| human glioblastoma cell line (U87-MG) | gelatin-based electrospun nanofibers | 220 μm | NR/2 days | anticancer therapies | (108) | |
| human breast tumor cells (LCC6/Her-2) | alginate | 250 μm | NR/4 days | anticancer therapies | (109) | |
| human breast cancer cell lines (BT49 and T47D) | basement membrane extract (BME) | 120 μm/125 μm | 11 spheroids/1 day | drug screening | (110) | |
| HT29 human colon carcinoma | 200–550 μm | ∼10000 spheroids/10 day | drug screening | (111) | ||
| human alveolar basal adenocarcinoma cell line (A549) | NR | 16 spheroids/72 h | 3D cell culture | (112) | ||
| human breast cancer stem cells (BCSCs) | Matrigel | NR | NR/5 days | tissue engineering | (113) | |
| acoustic | MCF-7/A549/human ovarian cancer cell line (A2780)/murine embryonic carcinoma cell line (P19) | NR | 6000 spheroids/24 h | 3D cell culture | (114) | |
| murine endothelial cell line (2H11)/ NIH 3T3/human embryonal kidney cell line (293FT) | 185.2 ± 50 μm | NR/9 h | tissue engineering | (115) | ||
| dielectrophoresis | human T lymphocyte cells (Jurkat)/mouse stromal cells (AC3) | at least 100 μm | NR/5 min | 3D cell culture | (116) | |
| human hepatoma cell line (HuH7) | 50–100 μm | NR/45 min | 3D cell culture | (117) |
a
NR: not reported.