Abstract
Purpose: :
The retrolental tissue in eyes with persistent fetal vasculature (PFV, persistent hyperplastic primary vitreous) is a vascularized, poorly differentiated tissue, which may harbor neuronal and endothelial progenitor cells. We have studied the potential of these cells to develop into either neuronal or endothelial elements in co–culture condition.
Methods: :
Retrolental membranes from eyes of patients with PFV were collected, subjected to collagen digestion. Cells were isolated and grown in serum enriched media. Cells were labeled with PKH67 red fluorescent cell linker, and transplanted onto retina explants derived from embryonic E8 chicks. Retinal explants were incubated between day 1 and day 8. Explants were collected, cryosectioned and immunostained for vimentin, neuronal/glial markers (nestin, NF–h, NF–m, GFAP), endothelial markers (CD34, AC133, CD31, cadherin–5 and VEGF–R2) and bone–marrow marker, CD45.
Results: :
Strong vimentin expression was detected in PFV cells in co–culture throughout whole time course. In co–culture condition, these cells proliferated and penetrated the immature chick retina. These cells did not express any detectable endothelial or neuronal markers between days 1–3. On day 3/4, CD31 (PECAM) expressing cells were detected within the transplanted cells. Between days 3–5, NF–m+ expressing cells from the host retina extended axonal–like sprouts into the transplants.
Conclusions: :
PFV cells are a undifferentiated self–replicating cell line. When transplanted into chick retina, they penetrate the host tissue and express CD31, an endothelial marker. These cells induce host retina to sprout axonal elements that penetrate them.
Keywords: retinal culture • vascular cells • differentiation