CORNEAL PRESERVATION &

STORAGE
DR.REKHA.
Lions Eye hospital
BANGALORE
IMPORTANCE OF STORAGE

SUCCESS OF PENENTRATING
KERATOPLASTY
DEPEND UPON CORNEAL
ENDOTHELIAL INTEGRITY.
CORNEAL STORAGE SYSTEM
AIM
 To maintain the endothelial viability &
integrity.
 Prevent corneal autolysis during storage
 Long term corneal storage.
 HISTORY
 First corneal preservation.

 1911 Mgitot in J.A.M.A.

 Stored cornea in Hemolyzed blood serum at

5-7deg.centigrade.

 Cornea viable for2days

 STORAGE TYPES
SHORT-4deg.C(days)
 Moist chamber(1day),M.K.Media(2-4days)

INTERMEDIATE-14days- 4deg.C(wks)
 K-Sol, Dexsol,OptisolGS,Procell, Eusol-C
LONG(months)
 EuropeanOrgan culture-120days(37deg.C)
 Eurosol-31deg.C-28days(New)
 Cryoprev.-Unlimited-(-80degC)
 MOIST CHAMBER METHOD
 Filatov &Castroviejo.
 Whole globe is stored in a sterile jar.
 Jar has saline Humidification.
 Temp of 4degrees C.
 Popular untill 1970.
 Simple & in- expensive.
 Grafting on emergency basis.
 McCAREY&KUFMAN
STORAGE
Corneo-scleral excision is necessary&
immersed with endothelial side up.
 Contains
Tc199,dextran-40 1%, PH7.0-7.5,Osmolality
295=355 & gentamycin sulphate75-150
micro gm/ml ,HEPES as buffer.
Phenol red as indicator.
 Scheduling of surgery
 STORAGE MEDIA
K-SOL: Tc 199,MEM&Earles media,
HEPES,Gentamicin,Chondroitin
sulphate2.5% .
1988 proprionibacteria contamination

DEXOL: MEM, 1.35%Chondrotin

Sulphate, 1mM Sodium pyruvate,
1mM non- essential aminoacids,
Antioxidants, 1%dextran40.
 OPTISOL.G.S.
 Introduced in 1991
 Contents
 MEM, 1.35%Chondrotin
Sulphate, 1mMSodiumpyruvate,1mM
non-essential aminoacids, Antioxidants,
1%dextran40,ATP,Iron,cholestrol,L-
hydroxyproline,Vitamine,2antibiotics-
Genticyn,Stepomycin. Expensive
 EUSOL-C
 Store at 4deg.C
 Storage time 14days, SIMILAR TO DEXOL
 CONTENTS
 Dextran,Sodium Piruvate,Glucose,Essential
&non-essential aminoacids, mineral salts,
Vitamins, Gentamin, hepesbuffer,
Bicarbonate, Phenol Red.

 FUNCTIONS OF CONTENTS
BASE MEDIA
a)Tissue-culture199-
aminoacids,salts,buffer&energy to support
life.
b)Minimum Essential Media(MEM) with
Earles salt- matches natural salts &buffers
of human tissue,electrolites similar to
aqueous
 FUNCTIONS OF CONTENTS
BUFFER
HEPES(N-2-hydroxyethyl-piperazine-N-
2ethane-sulfonic acid)
-used for cultivation of sensitive mammalian
cells
-provides optimal physiological Ph in the
range7-7.2
 COMMON FEATURES
OF MEDIA
ANTIBIOTIC
Gentamicin only /&Streptomycin

DEXTRAN
Polysaccharide,negatively charged.
Prevents tissue swelling.
Used alone does not provide sufficient tissue
stability & viability for extended storage.
 COMMON FEATURES
OF MEDIA
CHONDROTIN SULFATE
Polysaccharide,negatively charged
Endothelial integrity&acts as a osmotic
agent.Superior to dextran.Longer storage
time ENRICHED
Various salts,aminoacids essential &non-
essential,vit.,phosphates &antioxidents to
enhance cell & tissue viability & health during
storage.
COMMON MEDIA FEATURES
ATP precursors--> Provide energy for
pumping function.
Anti-Oxidents--->Neutralize metabolic
waste,maintain DNS synthesis
Vitamins-->Provides additional nutritional
cell supplements.
Sodium Piruvate&Glucose--> Energy supply
Colour Phenol Red-->Visual aid for pH
indication.
 CHECK MEDIA
The cornea which is stored in
MKmedia,Dexol,optisolGS.Thefollowing
parameters to be checked before using
Intact seal,Expiry date,Turbity, Colour (rose
red) ,precipitates & FB.
COLOUR CHANGE INDICATES
Yellow-- Bacterial contamination.
Red--- Unacceptable Ph.
Cloudy-- Contamination
 Ph COLOUR REFERENCE
CHART
 7.00

7.25

7.50

contimanation
 PROCELL

MEM,1.35%chondrotin sulphate,1mM
sodium pyruvate1mM,Non-essential
aminoacids,Antioxidants,Dextran40,
Humen insulin10mic.g/ml & Human
epidermal growth factor(hEGF10ng/ml) to
improve long term endothelial survival after
PKP .(experiments have not proved it, opth1994 feb101(2)252-9 LassJH et all)
 ORGAN CULTURE
 1936,by Archer & Trevor-Roper.
 Being used since1974.
 Refrigeration not required.
 Complicated,expensive & well trained
microbiologist.
EUROPEAN ORGAN CULTURE
31-37deg.C, EMEM, MEM, IMDM ,
HEPES, NaHCO3,Peni,Strep,Ampo-B,
BOVINE SERUM 2-10ML,Dextran4-10%
 ORGAN CULTURE
contents
Eagles media,Earles salt with out L-
glutamine,L-glutamine1%final
conc.,Decomlemented calf serum10%final
conc.Penicilllin 100units/ml,,
Gentamicin100microgram/ml.
AmphotericianB 0.25% microgram/ml.
 TISSUE CULTURE MEDIA
EUROSOL
New
Duration of storage 28days.
Temp 31deg.C.
Better then the European culture media.
Maintains intact endothelium, epithelium
& Keratocyte.
 ORGAN CULTURE
ADVANTAGES:Endothelial integrity
120days,Epithelial integrity35days,Culture
report prior to transplantation,stored at
4deg,centi.

DISADVANTAGES:Cornea thickens with

time,Microbial contamination, Complicated
expensive techniques, Greater loss of
endothelial afterPK.
 CRYOPRESERVATION
 Kaufman &Capella.1954
 Cornea is stored in Liquid Nitrogen with
DimethylSulfoxide(DMSO).
 DMSO,Prevents intracellular damage by ice
crystals.
 Not a procedure used.Holds a lot of
research interest.
 CRYOPRESERVATION
ADVANTAGES:Long-term storage at-
80deg.C.
DISADVANTAGES:Technically
difficult,Limited tissue tolerance to freez-
thaw. Rapid thaw at 37 deg.C.
 1964
 18months
 Tissue strored in Liquid nitrogen at --197deg C
 At the time of thawing the cial containing cornea is placed
in water bath at temp of 60dec C at 1min.
 Trained techiques for Freezing & thawing
 GLYCERINE PRESERVATION

Patch graft or Lamellar Keraoplasty.

Corneo-scleral button.
100%glycerine.
Enothelium is nonviable
Use with in 1 year.
Stored at room temp.
NOTE

 HYPOTHERMIC (CS+Hyperkalaemic)
 Storage 2wks,epithelial not well preserved,not
used

 ORGAN CULTURE
 At one time predominating in Europe
 Organ culture storage-4wks

 CRYOPRESERVATION
 Not used, tissure strored for emergency use
 FUTURE
 Improve endothelial cell layer viability
&compact stroma.
 Maintain Keratocyte viability.
 Reduce corneal autolysis during storage.
 Thin cornea in spite of long time storage.
 No contamenation
 Easy to use & transport
 Inexpensive.
Thank you