Hi, guys, I have a queation, please help me.
I used air liquid interface method to do 3D culture. This is the first time to do it.
the collagen gel prepared according to literature, 8 vol of collagen with 1 vol of 10X medium, then mix with 1 vol of reconsitution buffer.
I used 24 well plate, put an insert ( Millicell, cat. PICM01250) into one well, I added 300 ul of collagen gel on the bottom of the insert, put in the incubater for 30 min, after solid, added 350 ul collagen gel mix with cells on the top layer, outside of the insert, I added 450 ul medium, which did not above the up layer of collagen (this also mentioned by the ALI method), after 2days in the incubater, the up layer of the collagen evaporated some, the height of the collagen gel decreased a lot. I think this is not the normal situation, I don't know which step was wrong, please give me some suggestion.
Hi Letian,
I read one paper about the kinetics of the contraction of Collagen gel, for me, with more cells will contract fast.
for Skin model, the Collagen gel should not exposure to air directly, so it should not be evaporated, but i do not know which Skin model do you use. so sorry i can not help you.
Dear Yin Xiao & Letian Zhang,
I have experience in the in vitro 3D-skin model using our ovine collagen type I using ALI method. The contraction degree would be happen depending on the cells number mixed or seeded on the collagen gel or hydrogel. It is seldom that the hydrogel or gel would be dried up following time even though you are using ALI approach. However, at this point, I would rather ask you to evaluate the quality of your gel/hydrogel quality for long term culture even in submerged or ALI technique before you proceed to do your further experiment.
You could email me if you have further discussion. Thanks.
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