Cell growth in vitro directed by handmade patterns.
In this report, we show how the in vitro model of mechanically injured confluent monolayers of cultured mammalian cells, consisting in denudation by gentle scraping of areas in the monolayer, can be extended to obtain patterned cell cultures without using preadded attaching matrices. This work was done with a sinusoidal endothelial liver cell line. Patterns for cell growth were drawn in confluent monolayers by cell detaching with the aid of pipette tips followed by reincubation of the culture. In one or some d, acellular patterns were closed by cell migration and proliferation. For unveiling the pattern formed by migration and cell duplication, an enzymatic digestion with trypsin-collagenase solution was applied; after some min, only the migrating and younger cells filling the previous acellular pattern remained attached to the dish, and the now cellular pattern was clearly depicted. After stopping and recovering from the enzymatic treatment, the culture was ready for starting studies such as those related to migration, proliferation, cell-cell interactions. This method allows us to create simple and complex patterns, does not require preparation of the dishes with attaching matrices, and extracellular matrices in acellular areas are absent because of the enzymatic treatment, thus, potentially having many applications in cell culture techniques.
Reyes
Miguel R
MR
Laboratory of Cell Biology, Food and Nutrition Research Center, Faculty of Medicine, Universidad Juarez del Estado de Durango, Avenue Universidad S/N, Durango, Durango CP 34000, Mexico. mareyes@linux.ujed.mx
Lazalde
Brissia
B
D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans
Mol Biol Cell. 2001 Apr;12(4):831-45
11294890
Exp Biol Med (Maywood). 2002 Dec;227(11):1006-16
12486211
Hepatology. 2001 Jan;33(1):74-80
11124823
Dig Dis Sci. 1998 Mar;43(3):476-84
9539640
Mol Biol Cell. 1998 Apr;9(4):749-57
9529375
Science. 1997 May 30;276(5317):1425-8
9162012
In Vitro Cell Dev Biol Anim. 1999 Sep;35(8):441-8
10501083
Invest Ophthalmol Vis Sci. 2004 Apr;45(4):1088-94
15037573
Bone. 2002 Aug;31(2):288-95
12151081