Expression of a2, a5 and a6 subunits of integrin  in de-differentiated NIH3T3 cells by cell-free extract  of embryonic stem cells.
Generation of patient specific stem cells is among the ultimate goals in regenerative medicine. Such a cell needs to be functional when it transplants. Interaction between the matrix proteins and integrin adjust many cells' function such as adhesion, migration, cell cycle and self renewal in stem cells. In this study, NIH3T3 cells were dedifferentiated by mouse Embryonic Stem Cell (mESC) extract. The expression of pluripotency markers as well as a2, a5 and a6 integrin subunits were determined. NIH3T3 cells treated with mESC extract showed noticeable changes in cell morphology as early as day 2 post-treatment forming colonies similar to typical mESC morphology by day 8, after three passages. Alkaline phosphatase (ALP) assay and immunocytochemistry staining were performed for the induced reprogrammed cells. The results indicated that these colonies showed the ALP activity and they express Sox2 and Nanog. RT-PCR revealed that the colonies also express Oct3/4. NIH3T3 cells, ESC and reprogrammed cells expressed a2 integrin. a5 integrin expression was greatest in reprogrammed cells followed by the expression of this integrin in NIH3T3 which in turn was more than in ESC. a6A integrin was expressed in NIH3T3 cells while a6B integrin was expressed in ESC and in very low quantity was expressed in reprogrammed cells. These data provide evidence for both the generation of ES like cells from differentiated somatic cells and the expression profile of integrins after de-differentiation by mESC extract.
Biochemistry Department, Shiraz University of Medical Sciences, Medical School, Shiraz, Iran. zmostafavipour88@yahoo.co.uk
Zohreh
Z
Mostafavi-Pour
Sarva
S
Keihani
Tahereh
T
Talaei-Khozani
Pooneh
P
Mokaram
Majid
M
Fardaei
Leili
L
Rohani
Saeedeh
S
Ebadat
Ahmadreza
A
Sardarian
D000469:Alkaline Phosphatase; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D054337:Cell Dedifferentiation; D002454:Cell Differentiation; D002460:Cell Line; D002474:Cell-Free System / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000096:biosynthesis; D039421:Integrin alpha2 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D039482:Integrin alpha5 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D039503:Integrin alpha6 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis
D018398:Homeodomain Proteins; D039421:Integrin alpha2; D039482:Integrin alpha5; D039503:Integrin alpha6; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D000469:Alkaline Phosphatase