Organ Culture - Gastrointestinal biopsy (Dec/23/2007 )
hello,
I was looking in for any protocols for culturing gastrointestinal biopsy tissues, I will be using the tissues for virus culture, hence will also have to monitor the viability of the cells, so as to rule out false results of viral replication.
Would like to have a protocol or any books that would give me a detailed method of doing the procedure.
thekid
-thekid-
QUOTE (thekid @ Dec 23 2007, 10:11 AM)
hello,
I was looking in for any protocols for culturing gastrointestinal biopsy tissues, I will be using the tissues for virus culture, hence will also have to monitor the viability of the cells, so as to rule out false results of viral replication.
Would like to have a protocol or any books that would give me a detailed method of doing the procedure.
thekid
I was looking in for any protocols for culturing gastrointestinal biopsy tissues, I will be using the tissues for virus culture, hence will also have to monitor the viability of the cells, so as to rule out false results of viral replication.
Would like to have a protocol or any books that would give me a detailed method of doing the procedure.
thekid
if predominantly epithelial short-term culturing (~2-3h) may be successful in O2-satisfied Ringer solution;
-The Bearer-
Hello,
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
-thekid-
QUOTE (thekid @ Dec 25 2007, 05:25 PM)
Hello,
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
my first reply was rather conservative as normal gastric or intestinal epithelial cells are very difficult to culture; moreover, gastro-intestinal biopsies are hetergeneously composed of various tissues (epithelium, connective tissue, muscle), and may not meet a general protocol of culturing; in any case offer a coating, f.i. Matrigel;
you will succeed if your aim is to cultivate transfomerd cells which have reduced rates of anoikis;
-The Bearer-
QUOTE (The Bearer @ Dec 26 2007, 12:35 PM)
QUOTE (thekid @ Dec 25 2007, 05:25 PM)
Hello,
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
my first reply was rather conservative as normal gastric or intestinal epithelial cells are very difficult to culture; moreover, gastro-intestinal biopsies are hetergeneously composed of various tissues (epithelium, connective tissue, muscle), and may not meet a general protocol of culturing; in any case offer a coating, f.i. Matrigel;
you will succeed if your aim is to cultivate transfomerd cells which have reduced rates of anoikis;
if you are interested in intestinal epithelial cells they need excellent conditions to be cultured (beside if you only interested in oncogenic transformed cells); Matrigel is an ECM containing various ECM proteins; you have to coat your wells/plates of culture dishes before inoculation/seeding...
-The Bearer-
QUOTE (The Bearer @ Jan 5 2008, 09:57 PM)
QUOTE (The Bearer @ Dec 26 2007, 12:35 PM)
QUOTE (thekid @ Dec 25 2007, 05:25 PM)
Hello,
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
my first reply was rather conservative as normal gastric or intestinal epithelial cells are very difficult to culture; moreover, gastro-intestinal biopsies are hetergeneously composed of various tissues (epithelium, connective tissue, muscle), and may not meet a general protocol of culturing; in any case offer a coating, f.i. Matrigel;
you will succeed if your aim is to cultivate transfomerd cells which have reduced rates of anoikis;
if you are interested in intestinal epithelial cells they need excellent conditions to be cultured (beside if you only interested in oncogenic transformed cells); Matrigel is an ECM containing various ECM proteins; you have to coat your wells/plates of culture dishes before inoculation/seeding...
Is it necessary to use Matrigel during cultivation of primary gastric epithelial cells? Some scientist told me that usually coating with collagen will be enough.
-Roman80-
QUOTE (Roman80 @ Apr 15 2008, 09:34 AM)
QUOTE (The Bearer @ Jan 5 2008, 09:57 PM)
QUOTE (The Bearer @ Dec 26 2007, 12:35 PM)
QUOTE (thekid @ Dec 25 2007, 05:25 PM)
Hello,
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
Thanx for the suggestion, but I was hoping to use MEM and a CO2 incubator, I did read an article that the cells can survive up to 3 weeks if properly cared for.
Would you suggest the MEM and CO2, and I will be trying to grow more of the epithelial cells.
thekid
my first reply was rather conservative as normal gastric or intestinal epithelial cells are very difficult to culture; moreover, gastro-intestinal biopsies are hetergeneously composed of various tissues (epithelium, connective tissue, muscle), and may not meet a general protocol of culturing; in any case offer a coating, f.i. Matrigel;
you will succeed if your aim is to cultivate transfomerd cells which have reduced rates of anoikis;
if you are interested in intestinal epithelial cells they need excellent conditions to be cultured (beside if you only interested in oncogenic transformed cells); Matrigel is an ECM containing various ECM proteins; you have to coat your wells/plates of culture dishes before inoculation/seeding...
Is it necessary to use Matrigel during cultivation of primary gastric epithelial cells? Some scientist told me that usually coating with collagen will be enough.
from the late 80íes to the early 90ies, much efforts were done to improve primary culture of gastric epithelial cells; most efficient was Matrigel, collagen and poly-lysine coatings were less efficient;
if the those scientists have only general experience with other types of cells but never worked with gastric epithelial cells, be careful
-The Bearer-