Role of integrin beta1-like protein in proliferation and differentiation of cultured stem cells from midgut of Heliothis virescens.
Cultured midgut cells from Heliothis virescens larvae were incubated with anti-human integrin beta1 made in rabbit and then passed over a column of magnetic beads bound to anti-rabbit IgG (MACS, Miltenyi Bergisch Gladbach, Germany). Cells bound to integrin beta1 antibody also bound to the anti-rabbit IgG on the magnetic beads (MACS) and were retained in the column while it remained in the magnetic field. Non-bound cells were eluted at this time. They did not stain with anti-integrin antibody just after elution. Removing the column from the magnetic field allowed cells bound to the beads-integrin beta1 antibody to be eluted. All of these cells stained with human anti-integrin beta1 upon elution. Each cell fraction was cultured in medium for 3 days. During this time, the populations of cells tended to return to heterogeneous staining patterns characteristic of control populations. However, cells that did not stain immediately with anti-integrin beta1 antibody exhibited double the rate of multiplication and 8 times more differentiation than the integrin-antibody positive cells that eluted later, as well as the non-treated control cells. In a second experiment, midgut cells were incubated for 4 days with various titers of human anti-integrin beta1 to block surface integrin beta1-like reactive sites. Stem cells blocked with anti-integrin beta1 antibody during incubation exhibited double the rate of differentiation than non-treated control cells and those showing anti-integrin beta1-positive stain upon elution.
Loeb
Marcia J
MJ
Insect Biocontrol Laboratory, U.S. Department of Agriculture, Beltsville, Maryland, USA. geo.mar2@verizon.net
D000818:Animals; D000906:Antibodies; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D041981:Gastrointestinal Tract / Q000166:cytology*; D019476:Insect Proteins / Q000378:metabolism*; D019012:Integrin beta1 / Q000378:metabolism*; D007814:Larva / Q000166:cytology / Q000378:metabolism; D009036:Moths / Q000166:cytology / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
D000906:Antibodies; D019476:Insect Proteins; D019012:Integrin beta1