Abstract
The organoid culture method using completely isolated cells from fetal organ is a useful tool for studying organogenesis. Using this method, we investigated the renal development, especially renal tubulogenesis, by a combination of electronmicroscopy, lectin-histochemistry, and reverse transcription linked polymerase chain reaction (RT-PCR). Completely isolated cells from fetal mouse kidneys were seeded onto a membrane filter. After 7 days in culture, a dome-shaped cellular mass consisted of tubule-like structures was formed. Some short microvilli and basement membrane were observed. After 14 days in culture, microvilli became longer, and the number of microvilli was increased. The tubule-like structure showed a positive staining of lectins[Dolicos biflorus agglutinin (DBA) and Wheat germ agglutinin (WGA)]. Expression of aquaporin-2 (AQP-2) could be detected after 1 day in culture. When the culture periods were prolonged, the expression of AQP-2 was not detectable. Based on the findings of morphological features and lectin-histochemical staining patterns, we concluded that the tubule-like structure was recognized as a primitive collecting duct. Organoid culture method presented here is a useful tool for studying the development of the kidney.
