Adherence Capability of Cultivated Retinoblastoma Cells

Abstract

 

Purpose

 

To investigate adherence capability of cultivated retinoblastoma (RB) tumorspheres with prolonged cultivation.

 

Methods

 

RB cells from two Iranian patients were cultivated in DMEM supplemented with 15% FBS for four weeks in each passage. Fresh medium was added on a weekly basis and immunocytochemistry for synaptophysin was performed. All the experiments were done in duplicate. Cell attachment capability was studied during three consecutive passages.

 

Results

 

A biphasic population of cultivated cells was observed during the first week in each passage; composed of RB tumorspheres and a single-cell suspension overlying a layer of fibroblastic cells that had adhered to the bottom of flask. Early adherence of RB tumorspheres to the bottom of flask, while surrounded by fibroblasts, was observed in the second week and increased by the fourth week (figure).

 

Conclusions

 

Compared to previous data, this study demonstrated the adherence capability of RB tumorspheres to the underlying surface with prolonged cultivation; which after 4 weeks seems to be independent of the fibroblasts.  

 

Figure: Cultivated retinoblastoma (RB) cells. A-B: Note the presence of RB tumorspheres (circle 1), single-cell suspension (circle 2) in the first week C-F: Cultivated RB cells in the second week: the circled area shows adhered RB tumorspheres; merged FITC synaptophysine expression and DAPI in cultivated RB cells (C), adhered RB tumorsphere (D), DAPI of nucleus of RB and fibroblast cells (E), merged FITC synaptophysine expression and DAPI in cultivated RB cells (F).