Table 2.
Application of CR to generate primary cell cultures.
| Tissue origin | Case | Finding | Application | Ref. | |||
|---|---|---|---|---|---|---|---|
| Prostate | Matched human normal/tumor tissues (radical prostectomy) | 1 | CR normal and tumor cells are successfully established and characterized, maintaining low levels of differentiation in vitro. | In vitro and in vivo prostate cancer model | 100 | ||
| Matched human normal/tumor tissues (radical prostectomy) | 1 | Strigolactone analogues selectively kill CR tumor cells via inducing cellular stress and apoptosis. | Preclinical drug evaluation | 101 | |||
| Myc-driven mouse prostate tumor tissue (Hi-Myc transgenic C57BL/6 mouse model) | Not mentioned | CR prostate cancer cells from transgenic mice with Myc-driven prostate tumor are successfully cultured with tumorigenic ability. | Establishment of a Myc-driven prostate cancer model | 102 | |||
| Breast (male) | Human tumor tissue (freshly resected) | 1 | CR male breast cancer cells are successfully established and characterized. | In vitro model of male breast cancer | 103 | ||
| Breast (female) | Human tumor tissue (freshly resected) | Not mentioned | CR breast cancer cells are successfully established and characterized. | In vitro breast cancer model | 104 | ||
| Human tumor tissue (freshly resected) | 6 | CR breast cancer cells at early passages maintain main genetic characteristics of primary tumors. | In vitro breast cancer model | 83 | |||
| Human normal mammary tissue (prophylactic surgery) | 4 | CR enables heterogeneous culture of primary mammary cells. | Establishment of mammary cell line | 105 | |||
| Human DCIS tumor tissue (lumpectomies and mastectomies) | 19 | CR DCIS cells are cultured for 2 months expressing both luminal and basal marker and maintaining tumor heterogeneity. | In vitro DCIS model | 106 | |||
| Human tumor tissue (needle biopsy) | 5 | CR luminal-B breast cancer cells are established in 3 of 5 tissues, demonstrating similar gene expression profile to primary tumors. The CR cells enable the evaluation of drug sensitivity of tamoxifen, adriamycin and docetaxel. | In vitro model of luminal-B breast cancer; drug sensitivity test | 107 | |||
| Mouse tumor tissue (genetically engineered mouse models of triple negative mammary cancer) | 4 | CR cells retain tumor heterogeneity and epithelial cell differentiation, which is better than other methods. | A model for triple negative mammary cancer | 108 | |||
| Lung | Human tumor tissue (freshly resected) | 12 | NSCLC tumor cells are cultured in only 1 case. Normal epithelia outgrow cancer cells in CR condition. | Establishment of NSCLC cell lines | 97 | ||
| Human pleural effusion | 1 | CR cells from EGFR-mutant lung cancers maintaining tumor heterogeneity help the understanding of rociletinib resistance. | In vitro model of EGFR-mutant lung cancer | 109 | |||
| Human tumor tissue (biopsy) | 3 | ||||||
| Human normal and tumor tissue (freshly resected) | 1 | CR cells from respiratory papilloma help identify vorinostat as a therapeutic agent. | Individualized treatment | 110 | |||
| Human tumor tissue (freshly resected) | 14 | CR NSCLC cells are established and characterized and are applied to drug sensitivity test. | Drug sensitivity test | 111 | |||
| Human tumor tissue (freshly resected) | 10 | CR NSCLC cells maintain intratumor heterogeneity of original tumor by >90%. | In vitro NSCLC model | 112 | |||
| Pig lung tissue (newborn CFTR+/+ and CFTR−/− piglet) | Not mentioned | CR alveolar epithelia expanded in vitro allow analysis of bioelectric properties and liquid transport. | Establishment of an in vitro pulmonary edema model | 113 | |||
| Respiratory tract | Human airway tissue (excess lung donor tissue) | 1 | With a phenotype of adult stem cell-like cells, CR tracheal epithelium forms the upper layer of the ciliary airway in a gas-liquid interface culture system. | Establishment of a tracheal epithelium cell line | 12 | ||
| Human airway tissue (brushing) | Heathy | 18 | CR enables rapid cell expansion, maintaining airway epithelial cell characteristics and disease-specific functions. | Establishment of a disease model | 114 | ||
| Asthma | 11 | ||||||
| Cystic fibrosis | 8 | ||||||
| Human bronchial tissue (explanted lung) | Normal | Not mentioned | CR bronchial epithelium has the ability to differentiate into the upper and lower respiratory tract in both air-liquid interface and reconstructed mouse lung. | Tissue engineering | 115 | ||
| Cystic fibrosis | |||||||
| Human nasal and/or bronchial tissue (freshly resected, nasal brushing or bronchoscopy) | Newborns/infants/toddler (0–2 years) | 9 | CR airway epithelium maintains phenotype of the source cells after several passages and the immune response of the airways. | Establishment of a model for early-life respiratory disorders | 116 | ||
| School age children (4–11 years) | 6 | ||||||
| Adolescent/adult donors | 8 | ||||||
| Normal nasal airway tissue (nasal brushing) | 2 | Targeted genetic editing of CR primary airway epithelial cells by CRISPR-Cas9 reveals pro-inflammatory role for MUC18. | Biological function study | 117 | |||
| Human normal bronchial tissue (bronchial biopsy) | 19 | CR bronchial epithelial cells show multipotent differentiation property. | Tissue engineering | 69 | |||
| Human endobronchial tissue (brushing and biopsy) | 132 | Human airway epithelial cells from both endobronchial brushings and biopsies can be cultured by CR, showing better efficiency than other methods. Cryopreserved biopsies can also be expanded. | Establishment of cell lines for cell therapy or tissue engineering | 118 | |||
| Human normal airway epithelium (airway endoscopy or lung resections) | Not mentioned | CR primary airway epithelial cells combining with lung fibroblasts culture in 3D collagen scaffolds transplant into a decellularized rabbit trachea. | Tissue engineering | 119 | |||
| Human normal nasal cells (brush or curettage) | Not mentioned | Human nasal epithelial cells are expanded under CR conditions and inoculated into spheroid cultures to produce three-dimensional spheroids, as a model to characterize CFTR activity. | Establishment of a cystic fibrosis-specific disease model | 120 | |||
| Human normal bronchial tissue (fiberoptic bronchoscopy) | 3 | CR bronchial cells rapidly proliferate, express comparable levels of CYPs and are sensitive to BaP induction. | Establishment of in vitro toxicity testing model | 121 | |||
| Cystic fibrosis and non-cystic fibrosis tissue (explanted lung) | 6 | CR condition is modified for long-term primary culture of bronchial basal cells which maintains multipotent differentiation activity and CFTR channel function. | Establishment of primary bronchial cells for basic research and drug screen | 122 | |||
| Cystic fibrosis and non-cystic fibrosis tissue (freshly or cryopreserved explanted lung) | 8 | CR enables primary bronchial epithelial cells growing with larger number of cells than conventional culture. CR cells are expanded for testing CFTR modulators in Ussing Chamber. | Establishment of primary cystic fibrosis cells for drug assessment | 123 | |||
| Pig normal tracheobronchial airway tissue (newborn piglets) | 1 | CR porcine airway epithelial cells are successfully cultured and used for setting up a differentiated culture model at the gas-liquid interface. | A model for physiologic and pathophysiologic study | 124 | |||
| Esophagus | Esophageal tissue from patients with eosinophilic esophagitis (biopsy) | 8 | CR pediatric human esophageal epithelial cells are successfully cultured and maintain differentiation property. | Establishment of patient-specific cells for tissue engineering | 125 | ||
| Esophageal tissue from children with eosinophilic esophagitis (biopsy) | 28 | Patient-derived esophageal epithelial cell lines are successfully established which show disease-specific function. | Establishment of patient-specific model | 126 | |||
| Cornea | Normal limbal tissue | Human | 3 | CR maintains stable proliferation of normal limbal cells, with stable karyotype and the ability to form structured spheres in 3D culture. CR limbal cells differently response to several drugs. | In vitro model for corneal toxicity assessment | 127 | |
| Rabbit | 2 | ||||||
| Pig | 1 | ||||||
| Pancreas | Tumor tissue (freshly resected) | 3 | CR pancreatic cancer cells carry mutations identical with primary tumor, which enables therapeutic drug screen and identification of ERCC3-MYC interactions as a target in pancreatic cancer. | Establishment of in vitro and in vivo models for drug screen and drug target identification | 128 | ||
| PDX tumor tissue (first passage) | 3 | ||||||
| Pig normal pancreatic tissue (newborn pig pancreata) | 1 | Pancreatic epithelial cells are expanded under CR conditions and have the characteristics of a ductal epithelium, which can differentiate into functional cells at the gas-liquid interface. | A model for studying pancreas physiology and mechanisms of bicarbonate secretion | 129 | |||
| Liver | Human liver tissue (freshly resected from patients with cirrhosis, hepatitis C, maple syrup urine disease, or citrullinemia type 1 disease) | 11 | Primary hepatocytes are grown from 6 out of 11 specimens under CR condition, which are genetically identical with original tissues and retain strong CYP3A4, 1A1 and 2C9 activities. | Long term culture of patient-derived primary hepatocytes | 130 | ||
| Human tumor tissue (freshly resected) | 20 | Primary hepatocellular carcinoma cells continuously expand under CR condition and express tumor-specific marker. | Establishment of an in vitro model for precision medicine | 131 | |||
| Gastrointestinal tract | Human tumor tissue (freshly resected) | 1 | CR colorectal cancer cells are used to evaluate effect of a drug candidate IDF-11774. | Establishment of an in vitro model for drug assessment | 132 | ||
| Mouse small intestine tissue and tumor (wide type, CFTR ΔF508 and ApcMin/+ C57BL/6 mice) | 9 | CR intestinal epithelial cells can be expanded in vitro for up to 3 months, maintaining the specific function of the intestinal epithelium after 3D culture. | Establishment of a model for study of intestinal disorder | 133 | |||
| Mesenteric gland tissue (SD rat) | Not mentioned | CR meibomian gland cells is expanded in vitro maintaining functional sodium, chloride, and potassium channels, and cotransporters activities. | Establishment of a primary meibomian gland cell model for studying ion channels | 134 | |||
| Uterus and vagina | Human normal cervical tissue (hysterectomy) | 1 | CR primary cervical epithelial cells are adult stem cell-like cells. | Establishment of primary cervical epithelium cell line | 12 | ||
| Human tumor tissue (freshly resected liver metastasis of cervical cancer) | 1 | A stable CR cell line of neuroendocrine cervical cancer is established using CR, which identifies MYC overexpression as the primary driver of cervical cancer. | Establishment of a cell line for studying disease pathobiology | 135 | |||
| Human normal tissue (vaginal repair surgery) | 3 | CR primary vaginal epithelial cells are used for evaluating immunomodulatory effect of Houttuynia cordata. | Drug evaluation | 136 | |||
| Bladder | Human tumor tissue (radical cystectomy or transurethral resection) | 8 | CR bladder cancer cell lines are successfully established which are used for drug sensitivity test. | Drug sensitivity test | 137 | ||
| Skin | Human skin biopsy | Not mentioned | CR keratinocytes are genetically edited by CRISPR/Cas9, showing an important role of NLRP1 inflammasome upon UV sensing. | A model for biological study | 138 | ||
| Horse scrotal and neck skin biopsy | 2 | Equine keratinocytes acquire adult stem cell characteristics under CR conditions. | Tissue engineering | 139 | |||
| Cochlea | Mouse solid otic spheres (freshly resected from mouse strains of prestin-CreER, CAG-Cre, Ai14-tdTomato, and prestin-YFP) | Not mentioned | CR hair cells are successfully established, which are capable of expressing mature hair cell genes and responding to hair cell cues. | A model for biological study | 140 | ||
| Oral cavity | PDX tumor; Human tumor tissue (freshly resected) |
6 | CR cells from ACC show a cancer stem cell population driven by NOTCH1 and SOX10, and identify MYB fusion and CD molecules as markers for authentication and purification. | Establishment of ACC cancer stem cell line | 141 | ||
| Human normal and tumor tissue (freshly resected or needle biopsy) | 9 | CR cells from mucoepidermoid and other salivary gland neoplasms enable 2D, 3D and xenograft formation, and help identify the allosteric AKT inhibitor MK2206 as potential therapeutic agent. | Model of salivary gland neoplasm; Drug sensitivity test | 142 | |||
| Fish lip tissue (adult Mozambique tilapia) | Not mentioned | CR can rapidly and selectively culture lip epithelial cells. | A model for mechanism study | 143 | |||
| Dog tumor tissue (canine ameloblastoma of dog) | 4 | CR primary cells carry HRAS mutation. | A model for studying RAS-driven cancer | 144 | |||
| Mouse oral mucosa (freshly resected from C57BL) | 1 | CR oral mucosa epithelial cells are successfully established for long-term expansion. | Establishment of cells for potential tissue engineering | 145 | |||
Note: ACC, adenoid cystic carcinoma; CFTR, cystic fibrosis transmembrane conductance regulator; CYP, cytochrome P450 enzyme; DCIS, ductal carcinoma in situ; NSCLC, non-small cell lung cancer; PDX, patient-derived xenograft.