Objective: We describe a new method of immortalizing amniocytes with chromosome abnormalities to generate renewable resources for studying rare diseases. Methods: Three methods were investigated by randomly dividing 180 cases of adherent amniocytes into groups A, B, and C. Group A cells were digested with 0.25% trypsin for 10, 20, 30, 60, 90, and 120 mins. Group B and C cells were digested with 0.25% trypsin for 3-5 minutes. Group A and B cells were then transfected with PT67 cell-produced

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... T. Group C digested cells underwent liposomemediated SV40LT-transfection. Results: The percentage of clones immortalized by SV40LT transfection in groups A, B, and C were 18.3, 5.0, and 16.7%, respectively, after G418 screening. Group A produced a higher percentage of immortalized clones than did groups B and C, but only significantly differed from group B. The karyotypes and chromosome 13, 18, 21, X, and Y fluorescence signals in transfected cells of all groups were identical to those of the primary cells after passaging for 10-15 generations. Conclusions: The trypsin-mediated SV40LT transfection used in group A can be applied for routine establishment of amniocyte lines to obtain a renewable resource for the study of rare diseases and as quality control materials for prenatal diagnosis.