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Comment
. 2009 Jun;13(6):1186-7.
doi: 10.1111/j.1582-4934.2008.00651.x. Epub 2009 Jan 14.

Avoiding lentiviral transduction culture induced MSC senescence

Yung-Wei Pan, Peter Kurre
Comment

Avoiding lentiviral transduction culture induced MSC senescence

Yung-Wei Pan et al. J Cell Mol Med. 2009 Jun.
No abstract available

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Figures

Figure 1
Figure 1
(A) Lentiviral transduction of non-adherent murine mesenchymal stem cells (MSC). MSC were exposed to VSV-G pseudotyped green fluorescent protein (GFP) expressing lentivirus for 1 hr at the indicated multiplicities of infection (MOI), alternatively in adherence after plating 16–24 hrs earlier (open symbols), or in suspension (closed symbols). (B) Gene transfer to mMSC is improved under serum free conditions. MSC were thawed from liquid nitrogen, washed twice in phosphate buffered saline (PBS), resuspended in media, and immediately exposed to lentivector at the indicated MOI. All transductions were performed over 1 hr in 1ml final volume, in the presence of protamine sulphate at 4 μg/ml. At the end of transduction cells were washed twice in PBS and plated in Iscoves based MSC expansion medium, as described by Peister [7]. Retrovirus vector was produced as previously described [6]. For transduction in suspension, cells were trypsinized from log-phase expansion culture, or thawed from cryopreservation (5% dimethyl sulfoxide [DMSO]) in liquid nitrogen and washed in PBS. Averages from multiple repeat determinations are shown. Data were analysed using the paired two-tailed Student’s t-test. P-values of less than 0.05 were considered significant.
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