Background: Estimate the prevalence of vitamin D deficiency among patients diagnosed with chronic cholestasis and investigates the association between their clinical manifestations and lab results of 25(OH) D levels.Methods: A cross sectional study of 50 patients aged > 3 months up to 18 years, who had medical care in the Beni-Suef University hospital, Beni-Suef governorate, Egypt. All patients were fully investigated including routine labs in addition to 25- [OH] D levels and dual energy X-ray absorptiometry (DXA).Results: Mean age was 6.5±4.5 years. Mean serum 25 (OH) D levels was 37.9±28.2; 30% of patients had 25 (OH) D <20 ng/ml and 26% had 25 (OH) D 20-<30 ng/ml. Low BMD was diagnosed in 73.1% of patients. The spine DXA BMD ranged between -5.4 and -1.4 (-2.9±0.9 Z-scores). A delayed milestone was reported in 32% of patients, and 56% had signs of rickets.Conclusions: Deficiency of Vitamin D, signs of rickets and osteoporosis were evident in chronic cholestasis patients. No statistical significance was detected between 25- D [OH] levels and clinical, laboratory and radiological findings.