307) Each passaged miPS cells on these scaffolds were cryopreserved successfully and the revived cells showed high viability and proliferation. |
PMID:23166055 DOI:10.1002/term.1640 |
2015 Journal of tissue engineering and regenerative medicine |
* Hydrogels as feeder-free scaffolds for long-term self-renewal of mouse induced pluripotent stem cells. |
- Expanding undifferentiated induced pluripotent stem (iPS) cells in vitro is a basic requirement for application of iPS cells in both fundamental research and clinical regeneration. In this study, we intended to establish a simple, low cost and efficient method for the long-term self-renewal of mouse induced pluripotent stem (miPS) cells without using feeder-cells and adhesive proteins. Three scaffolds were selected for the long-term subculture of miPS cells over two months starting from passages 14 to 29: 1) a gelatin coated polystyrene (Gelatin-PS) that is a widely used scaffold for self-renewal of mouse embryonic stem (mES) cells; 2) a neutral hydrogel poly(N,N-dimethylacrylamide) (PDMAAm); and 3) a negatively charged hydrogel poly(2-acrylamido-2-methyl-propane sulfonic acid sodium salt) (PNaAMPS). Each passaged miPS cells on these scaffolds were cryopreserved successfully and the revived cells showed high viability and proliferation. The passaged miPS cells maintained a high undifferentiated state on all three scaffolds and a high level of pluripotency by expressing differentiation markers in vitro and forming teratomas in SCID mice with derivatives of all three germ layers. Compared to Gelatin-PS, the two hydrogels exhibited much better self-renewal performance in terms of high proliferation rate and level of expression of undifferentiated gene markers as well as efficiency in pluripotent teratoma formation. Furthermore, the PNaAMPS hydrogel demonstrated a slightly higher efficiency and simpler operation of cell expansion than the PDMAAm hydrogel. To conclude, PNaAMPS hydrogel is an excellent feeder-free scaffold because of its simplicity, low cost and high efficiency in expanding a large number of miPS cells in vitro. |
(1)80 *null* | (15)5 as | (29)3 exposed | (43)2 expanded |
(2)47 and | (16)5 expressed | (30)3 have | (44)2 expressing |
(3)39 were | (17)5 of | (31)3 isolated | (45)2 had |
(4)26 in | (18)5 which | (32)3 showed | (46)2 labeled |
(5)12 (MSCs) | (19)4 (ASCs) | (33)3 we | (47)2 may |
(6)11 was | (20)4 has | (34)2 (ASCs), | (48)2 play |
(7)10 to | (21)4 is | (35)2 (BMSCs) | (49)2 present |
(8)9 with | (22)4 on | (36)2 (ECFCs) | (50)2 remains |
(9)7 are | (23)4 seeded | (37)2 (HSCs) | (51)2 such |
(10)6 can | (24)4 within | (38)2 after | (52)2 the |
(11)6 for | (25)3 (ASC) | (39)2 by | (53)2 via |
(12)6 from | (26)3 (ECs) | (40)2 caused | |
(13)6 or | (27)3 (HUVECs) | (41)2 cultured | |
(14)6 that | (28)3 (hMSCs) | (42)2 derived |
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