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14163 Berlin
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immunologie@vetmed.fu-berlin.de
Introduction
The culture of primary kidney epithelial cells usually involves tissue digestion after nephrectomy. Urinederived epithelial cells (UECs) offer a non-invasive source of urinary tract epithelial cells. To optimize yield and purity, isolation methods for equine and canine UECs were compared.
Methods
Urine samples from dogs (n = 74) were collected from spontaneous midstream urine. Equine samples (n = 21) were collected during general anesthesia induction through a routine urinary catheter. All urine samples were examined by dipstick, microbiological culture, and microscopy for crystals and cell density.
Cells were seeded onto either gelatine- or Matrigel-coated (1:25) well plates and cultured at 37 °C and 5% CO₂ for 14 days. Medium exchange and microscopic examination were performed daily. Cells were passaged at 80–100% confluence until cryopreservation or characterization.
Results
UECs were successfully cultured from 28.6% of equine urine samples. Gelatine-coating showed a 37.5% and Matrigel-coating a 25.0% success rate. Canine UECs were cultured in 8.1% of cases, with gelatinecoating showing a 4.6% and Matrigel-coating a 9.6% success rate. Bacterial or fungal contamination terminated 24.3% of canine and 19.1% of equine cultures. The addition of 0.1% Y-27632, 0.2% primocin, and 15% FBS to the culture medium showed the most promising growth.
Conclusions
Isolation of equine UECs using gelatine-coated wells and medium containing 15% FBS and 0.1% Y-27632 shows promising results. Isolation of UECs from dogs remains challenging due to contamination and nongrowth, requiring further optimization.