Immortalizing Mesenchymal Stromal Cells from Aged Donors While Keeping Their Essential Features

Table 2

Variations of the transduction method assayed, detailing retrovirus employed, culture conditions for Phoenix cells, spinoculation conditions, and VPA concentration employed to induce transgene expression.

Experiment	Cells	Retrovirus	φNX-A culture	Spinoculation	VPA (mM)
1	MSC#1; MSC#2	SV40LT+hTERT	32°C 24 h	1000 × 60 min	0
2	MSC#1; MSC#3	(1) SV40LT+hTERT; (2) SV40LT; (3) hTERT	32°C 24 h	1000 × 60 min	0-0.5
3	MSC#4; MSC#5	SV40LT	32°C 24 h	1000 × 45 min	0.5-2
4	MSC#6	SV40LT	32°C 48 h	800 × 45 min	0.5-2
5	MSC#7; MSC#8	SV40LT	32°C 48 h	800 × 30 min	1-3
6	MSC#9; MSC#1; MSC#4; MSC#10	SV40LT	32°C 48 h	800 × 45 min	2
7	T-MSC#6; T-MSC#8; T-MSC#9; T-MSC#10	hTERT	32°C 48 h	800 × 45 min	2
8	MSC#11; MSC#12; MSC#13	SV40LT	32°C 48 h	800 × 45 min	0.5-5