| Publication number |
CLPUB00369 |
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| Authors |
Tao W., Stabila P.F., Dean B.J., Bell W. |
|---|
| Title |
Construction, characterization and safety testing of CNTF secreting mammalian cell lines for encapsulated cell therapy. |
|---|
| Citation |
Invest. Ophthalmol. Vis. Sci. 43:S2312-S2312(2002) |
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| Web pages |
https://iovs.arvojournals.org/article.aspx?articleid=2419568 |
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| Abstract |
Purpose: The objective of the current study was to construct,
characterize, and safety evaluate the CNTF secreting cell lines that will
be used for encapsulated cell therapy for treating retinal degenerative
diseases.
Methods: Two stable CNTF secreting human mammalian cell lines were
established by transfecting human RPE cells with a CNTF gene containing
plasmids. The CNTF secreting cell lines were designated NTC-201-10 and
NTC-201-6A. The growth characteristics, morphology and CNTF production were
monitored for over one year in culture. Their encapsulated performance was
evaluated both in vitro and in vivo: CNTF production was quantified by
ELISA, and bioactivity and specificity of CNTF were evaluated in a TF-1
CN5a.1 cell bioassay. Viral safety and tumorigenecity of the cell lines
were also examined.
Results: Both NTC-201-10 and NTC-201-6A cell lines maintained normal RPE
morphology with stable CNTF output in vitro for over a year. Un-
encapsulated NTC-201-10 cell line produced ~200 ng/106 cells/day, and
NTC-201-6A cell line produced ~800 ng/106 cells/day. Cell-secreted CNTF was
biologically active, with specific activity 2~3 fold higher than that
recombinant CNTF. Furthermore, the observed bioactivity was completely
blocked by a goat anti-CNTF neutralizing antibody. When encapsulated, both
cell lines secreted efficacious levels of CNTF in vivo, which was
quantified upon device explant at 1~5 ng/day for NTC-201-10 and 10~20
ng/day for NTC-201-6A, respectively. The cell lines tested negative for
viruses and are not tumorigenic in nude mice.
Conclusions: Two stable CNTF secreting cell lines were successfully
established which maintained normal growth characteristics and morphology
for over one year in vitro, and produced biologically active CNTF. Their
encapsulated performance and safety profile make them ideal for
encapsulated cell therapy.
|
|---|
| Cell lines |
CVCL_IQ64; ARPE-19/CNTF-6A CVCL_IQ63; ARPE-19/CNTF-10 |
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