Neonatal jaundice requires a differential diagnosis between unconjugated or indirect hyperbilirubinemia and direct or conjugated hyperbilirubinemia, characteristic of neonatal cholestasis, which is always pathological. It is recommended in all newborns of 15 days or older with jaundice to perform differential bilirubin to be able to detect it. Cholestasis is defined by direct bilirubin > 1 mg/dl (17 µmol/L) and its estimated frequency during the neonatal period is 1:2,500 live births. The most frequent etiologies are infectious, genetic/metabolic, obstructive, endocrinological, toxic, immunological. The most common cause is biliary atresia, which is clinically manifested by jaundice and acholia, so in patients with these signs that diagnosis should always be considered as the first option to rule out. An early diagnosis of neonatal cholestasis is essential to optimize management of treatable causes and improve prognosis. Unfortunately, diagnosis is not always early. That is why the Working Group of the Latin American Society of Pediatric Gastroenterology, Hepatology and Nutrition carried out this review of diagnosis, treatment and follow-up of neonatal cholestasis, whose objective is to provide a useful tool that allows early diagnosis and improves the prognosis of children with neonatal cholestasis.