. 2020 Jul 21;11:301. doi: 10.1186/s13287-020-01783-8

Table 3.

Summary of culture methods used in OMEC sheet preparation

Ref.	Culture system	Substrate	Feeder layer	Nutrient	Air lifting	% SC	Medium	GMP	Carrier	Culture time (days)
[13]	S	dAM	3T3	10% FBS	Y	–	DMEM/F12 (1:1)	N	dAM	14–21
[14]	S	CellSeed^b	3T3	NA	N	2.1 ± 0.9	NA	N	Carrier-free*	14
[15]	S	dAM	3T3	10% FBS/10% AS	Y	–	DMEM/F12 (1:1)	N	dAM	15–16
[16]	S	dAM	3T3	10% FBS^a	Y	–	DMEM/F12 (1:1)^a	N	dAM^a	14
[17]	S	dAM	3T3	5% AS	Y	–	KGM	N	dAM	15–16
[18]	S^a	dAM^a	3T3^a	U^d	Y^a	–	DMEM/F12 (1:1)^a	N^a	dAM	U^d
[19]	S	dAM	3T3	10% FBS	Y	–	DMEM/F12 (1:1)^a	N	dAM	> 14
[20]	S	dAM	3T3	5% FBS	N	–	SHEM	N	dAM	14–21^a
[21]	S	dAM	3T3	5% FBS	N	–	U	N	dAM	14–21^a
[22]	S	dAM	3T3	10% AS	N	2.0 ± 1.0	DMEM/F12 (1:1)	N	dAM	18–21
[23]	S	Fibrin^c/dAM	3T3	4% AS	Y	–	DMEM/F12 (1:1)	N	U	NA
[24]	S	dAM	3T3	5% Serum	Y	–	KGM	N	dAM^a	15–16
[25]	S^a	dAM	3T3	U^d	Y	–	DMEM/F12 (1:1)^a	N	dAM	15–16
[26]	S	CellSeed^b	3T3	NA	N	3.4 ± 2.06	NA	N	Carrier-free**	U^e
[27]	S	Fibrin^c/dAM	3T3	4% AS	Y	–	DMEM/F12 (1:1)	N	Fibrin group: carrier-free*** AM group: denuded AM	8–16 (Fibrin)/NA (dAM)
[28]	S	dAM	3T3	5% FBS	N	–	SHEM	N	dAM	14–21^a
[29]	S^a	dAM^a	3T3	10%^a FBS/5%^b AS	Y	–	U^d	Y	dAM^a	8–9
[30]	S^a	dAM^a	3T3	10%^a FBS/%^c AS	Y	–	U^d	Y	dAM^a	8–9
[31]	E	iAM	N	AS	U	~ 12	DMEM/F12 (3:1)	Y	iAM	21
[32]	S	dAM	3T3	10% FBS/10% AS	Y	–	DMEM/F12	N	dAM	7
[33]	S	dAM	N	Serum-free	N	–	KBM-2	N	dAM	14–21
[34]	S^a	dAM	3T3^a	10% FBS^a	Y	–	DMEM/F12 (1:1)^a	N	dAM	14–21
[35]	S	BM-free	3T3	10% FBS	N	NA	DMEM/F12 (3:1)	Y	Carrier-free****	7–12
[36]	S	dAM	3T3	5% FBS	Y	–	DMEM/F12 (3:1)	N	dAM	U^f

AM amniotic membrane, AS autologous serum, BM-free biomaterial-free, dAM denuded amniotic membrane, iAM intact amniotic membrane, E explant, FBS fetal bovine serum, DMEM/F12 Dulbecco modified Eagle’s medium (DMEM) with HAM F12 mixture, GMP good manufacturing practice, KGM keratinocyte growth medium, KBM-2 serum-free Keratinocyte Growth medium, N no, NA not available, S suspension, SHEM supplemented hormonal epithelial medium, U unclear, Y yes, 3T3 3T3 murine fibroblasts, %SC percentage of transplanted stem cells

^aAccording to the referenced protocol in the paper

^bCellSeed, temperature-responsive cell-culture inserts (CellSeed Inc., Tokyo, Japan)

^cFibrin-coated inserts

^dConflicting data among the referenced studies

^eFor at least 4 days after the confluence

^fFor at least 5 days after the confluence and then air-lifted for 1 to 2 days

*Supporter

**Polyvinylidene fluoride (PVDF) ring

***Filter paper

****Support mesh