Abstract
A recent remarkable study has shown that when mouse NIH-3T3 fibroblasts are exposed to an embryonic stem cell (ESC) extract, the majority of them expresses the Oct-4 gene, form ESC-like colonies, and embryoid-like bodies that differentiate into cells of the three germ layers. The use of cell extracts for inducing cell dedifferentiation could be a powerful system to obtain large quantities of pluripotent cells. It is thus of crucial importance that the robustness of this method of cell transdifferentiation is tested by other laboratories before it is advanced to a more ambitious use in cell therapy programs. We report here our experimental observations using the same reprogramming protocol on STO and NIH-3T3 mouse fibroblasts. Three are the main results: first, we confirmed an enduring reprogramming activity of the ESC extract, although on a much smaller number of cells that varies from ∼0.003 to 0.04% of the total population of fibroblasts and with an effect limited to the induction of Oct-4 and Rex-1 gene expression and alkaline phosphatase activity. Second, the expression of OCT-4, SSEA-1, and Forssman antigen proteins was never detected. Third, our work has clearly demonstrated that ESCs may survive the procedure of extract preparation, may be source of contamination that is expanded in culture and give false positive results.
| Original language | English |
|---|---|
| Pages (from-to) | 394-406 |
| Number of pages | 13 |
| Journal | Cloning and Stem Cells |
| Volume | 9 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Sep 1 2007 |
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ASJC Scopus subject areas
- Biotechnology
- Developmental Biology
- Applied Microbiology and Biotechnology
Cite this
Mouse fibroblasts are reprogrammed to Oct-4 and Rex-1 gene expression and alkaline phosphatase activity by embryonic stem cell extracts. / Neri, Tui; Monti, Manuela; Rebuzzini, Paola; Merico, Valeria; Garagna, Silvia; Redi, Carlo Alberto; Zuccotti, Maurizio.
In: Cloning and Stem Cells, Vol. 9, No. 3, 01.09.2007, p. 394-406.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mouse fibroblasts are reprogrammed to Oct-4 and Rex-1 gene expression and alkaline phosphatase activity by embryonic stem cell extracts
AU - Neri, Tui
AU - Monti, Manuela
AU - Rebuzzini, Paola
AU - Merico, Valeria
AU - Garagna, Silvia
AU - Redi, Carlo Alberto
AU - Zuccotti, Maurizio
PY - 2007/9/1
Y1 - 2007/9/1
N2 - A recent remarkable study has shown that when mouse NIH-3T3 fibroblasts are exposed to an embryonic stem cell (ESC) extract, the majority of them expresses the Oct-4 gene, form ESC-like colonies, and embryoid-like bodies that differentiate into cells of the three germ layers. The use of cell extracts for inducing cell dedifferentiation could be a powerful system to obtain large quantities of pluripotent cells. It is thus of crucial importance that the robustness of this method of cell transdifferentiation is tested by other laboratories before it is advanced to a more ambitious use in cell therapy programs. We report here our experimental observations using the same reprogramming protocol on STO and NIH-3T3 mouse fibroblasts. Three are the main results: first, we confirmed an enduring reprogramming activity of the ESC extract, although on a much smaller number of cells that varies from ∼0.003 to 0.04% of the total population of fibroblasts and with an effect limited to the induction of Oct-4 and Rex-1 gene expression and alkaline phosphatase activity. Second, the expression of OCT-4, SSEA-1, and Forssman antigen proteins was never detected. Third, our work has clearly demonstrated that ESCs may survive the procedure of extract preparation, may be source of contamination that is expanded in culture and give false positive results.
AB - A recent remarkable study has shown that when mouse NIH-3T3 fibroblasts are exposed to an embryonic stem cell (ESC) extract, the majority of them expresses the Oct-4 gene, form ESC-like colonies, and embryoid-like bodies that differentiate into cells of the three germ layers. The use of cell extracts for inducing cell dedifferentiation could be a powerful system to obtain large quantities of pluripotent cells. It is thus of crucial importance that the robustness of this method of cell transdifferentiation is tested by other laboratories before it is advanced to a more ambitious use in cell therapy programs. We report here our experimental observations using the same reprogramming protocol on STO and NIH-3T3 mouse fibroblasts. Three are the main results: first, we confirmed an enduring reprogramming activity of the ESC extract, although on a much smaller number of cells that varies from ∼0.003 to 0.04% of the total population of fibroblasts and with an effect limited to the induction of Oct-4 and Rex-1 gene expression and alkaline phosphatase activity. Second, the expression of OCT-4, SSEA-1, and Forssman antigen proteins was never detected. Third, our work has clearly demonstrated that ESCs may survive the procedure of extract preparation, may be source of contamination that is expanded in culture and give false positive results.
UR - http://www.scopus.com/inward/record.url?scp=38049086046&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=38049086046&partnerID=8YFLogxK
U2 - 10.1089/clo.2006.0011
DO - 10.1089/clo.2006.0011
M3 - Article
C2 - 17907950
AN - SCOPUS:38049086046
VL - 9
SP - 394
EP - 406
JO - Cloning and Stem Cells
JF - Cloning and Stem Cells
SN - 1536-2302
IS - 3
ER -