|
NR 6/2004
|
|
|
|
|
|
|
|
|
Hodowla nabłonka
rogówki – porównanie potencjału mitotycznego komórek uzyskanych
od dawców żywych i po ¶mierci
Culture of the corneal
epithelium – comparison of the mitotic potential of limbal cells
from living and cadaveric donors
Edward A. Wylęgała1, Dariusz
Dobrowolski1, Bożena Gabryel2, Andrzej
Małecki2,
Dorota Tarnawska1, Antonina Jurewicz1,
Jolanta Łogiewa-Toborek1
1Z Oddziału Okulistycznego Okręgowego Szpitala
Kolejowego w Katowicach
Ordynator: dr n. med. Edward Wylęgała
2Z Zakładu Farmakologii Katedry Farmakologii Wydziału
Lekarskiego ¦l±skiej Akademii Medycznej w Katowicach
Kierownik: dr hab. n. med. Andrzej Małecki |
|
|
| Summary: |
Purpose: We analyse
effectiveness of corneal limbal cells' culture prepared
from heart-beating organ donors, that include living
donors and from cadaver donors buttons following 3 days
storage in 4°C in Likorol.
Material and methods: For experiment 12 adults (living
and heart-beating organ donors) aged 28-63 (mean 46.3)
and 12 corneal buttons of cadaver (aged 18-51, mean
34.1) were qualified. Tissue samples (1 mm2) were taken
from superior corneal limbus. Sample from living donor
obtained during routine operation was sent immediately
to laboratory, as well as from heart-beating organ
donor. The limbal biopsy of preserved cornea was taken
after 3 days storage in 4°C (Likorol). Samples were
treated with trypsin / EDTA solution before culture.
Collected cells in similar density in 1 ml of medium
laid on dishes inserts, covered with fibrin (Tissucol)
and cultured in presence of feeder layer of fibroblasts
(L929 line). Epithelial cells were cultured for 14 days
at 37°C in humidified 5% CO2 atmosphere in supplemented
2:1 mixture of DMEM and Ham`s F12. On the 14th day cells
were collected. Number of cells per 1 ml of medium was
counted by cytometer. Immunostaining for epithelium type
(Keratin 3) was performed.
Results: The number of cells obtained from
cadaver donors reached 184.2±14.9% whereas from living
donors revealed 1013.1±104.2%, increase in relation to
number of delivered cells. We observed only 0.83±0.3
colonies per microscopic area in cultures from preserved
tissue versus 6.67±0.6 colonies in cultures from living
donor.
Conclusions: The preservation in 4°C in Likorol
significantly decreases proliferative potential of the
corneal limbus. |
| Słowa kluczowe: |
nabłonek rogówki, komórki
macierzyste r±bka, hodowla tkankowa. |
| Key words: |
corneal epithelium, limbal
stem cells, organ culture. |
|
|
|
|
|
|
