Novel biomanufacturing platform for large-scale and high-quality human T cells production

Table 1 Summary of key parameters in human T cell biomanufacturing

Biomanufacturing Parameters^a,b,c		Scale (mL)	VCD_max^d (10⁶ cells/mL)	TVC (10⁶ cells)	Viability^d (%)	μ^d(hr^− 1)	Td^c(hr)
Medium	AIM-V	30	1.90 ± 0.11	56.9 ± 3.2	76.8 ± 2.5^e	0.030 ± 0.001	23.2 ± 0.8
	OpTmizer	30	2.83 ± 0.12	84.8 ± 3.6	87.5 ± 1.4^e	0.030 ± 0.004	23.1 ± 2.9
	ImmunoCult	30	3.49 ± 0.52	104.6 ± 15.5	91.8 ± 1.1	0.041 ± 0.000	17.1 ± 0.2
Seed and stimulation	1st stimulated Day 0 seed	80	8.33 ± 0.11	666.0 ± 8.5	93.0 ± 1.4	0.053 ± 0.001	13.0 ± 0.2
	1st stimulated Day 2 seed	80	1.89 ± 0.04	151.2 ± 3.4	95.0 ± 1.4	0.029 ± 0.002	23.8 ± 1.5
	1st stimulated Day 5 seed	80	0.55 ± 0.02	44.1 ± 2.0	96.0 ± 1.4	0.007 ± 0.001	99.5 ± 17.2
	2nd stimulated Day 0 seed	80	3.98 ± 0.06	318.4 ± 4.5	93.0 ± 0.0	0.042 ± 0.002	16.7 ± 0.7
Scale-up^f	T75	10	4.05 ± 0.13	40.5 ± 1.3	90.5 ± 0.7	0.034 ± 0.002	20.4 ± 0.9
	SF125	30	3.65 ± 0.18	109.4 ± 5.3	91.5 ± 2.1	0.041 ± 0.001	16.9 ± 0.4
	Spinner	80	3.98 ± 0.06	318.4 ± 4.5	93.0 ± 0.0	0.042 ± 0.002	16.7 ± 0.7
	Bioreactor	800	6.40 ± 0.46	5120.0 ± 367.7	94.0 ± 1.4	0.046 ± 0.002	15.2 ± 0.7

Notes
^aThe human T cell biomanufacturing was performed in spinner flasks for 4 days with basal medium of OpTmizer, feed of 30 U/mL IL-2 and 1st stimulation on Day 0 and Day 2, unless otherwise specified.
^bSamples from multiple donors were used in the biomanufacturing process development, and T cells isolated from the same donor were used in the evaluation of one biomanufacturing parameter (i.e., medium, seed and stimulation, and scale-up). Significant difference between bioreactor runs in triplicate with p ≤ 0.05 was considered in the two-tailed t test. The results of cross donor comparison are presented in Fig. 3.
^cAll data are presented as mean ± standard error of the mean (SEM).
^dThe variation in cell growth parameters, such as VCD, viability and cell growth rate, was caused by the different biomanufacturing process parameters.
^eThe viabilities of the T cells when reaching maximal VCD are presented. The relatively low harvest viability in the medium evaluation study was caused by an extended seed culture process in the early stage of process development, i.e. 6 days instead of 4 days.
^fThe scale-up strategy was designed following the procedure described in Fig. 1.

ISSN: 1754-1611