Derivation and Culture of Putative Parthenogenetic Embryonic Stem Cells in New Gelatin Substrates Modified with Galactomannan

다국어 초록 (Multilingual Abstract)

Human embryonic stem cells (ESC) lines to be used for cell therapies must be created and maintainedunder strict conditions, excluding the use of undefined supplements. Two key steps in the creation of a new embryonicstem cell line are adherence to the substrate and derivation towards the formation of a primary colony. Thebovine parthenote embryo model was used to test different matrices of gelatin nanofibers and gelatin/galactomannanfilms to be used for ESC derivation and culturing. Gelatin/galactomannan films were made in two concentrations ofgalactomannan, 0.1 and 0.3%, in an aqueous solution of gelatin and tested for gel cytotoxicity using cumulus cells(CCs). CCs showed normal cell morphology, with no sign of lysis or degeneration in any of the matrices tested. Innercell masses of parthenote blastocysts (n=116) were placed onto the gel matrices for culture. There were three or fourrepeats for each matrix. Our results showed a good rate of inner cell mass (ICM) adherence on the gelatin/galactomannanfilms (41%-44%) and one derivative of the gel nanofiber (17% adherence to the substrate). These resultsencouraged us to try new gelatin formulations to increase the rates of derivation and cell proliferation under definedculture conditions to comply with good manufacturing practice directives for the potential therapeutic use of ESCs.

참고문헌 (Reference)

1 J. A. Thomson, 282 : 1145-, 1998

2 N. Desai, 30 : 609-, 2013

3 H. Darr, 133 : 1193-, 2006

4 K. Gauthaman, 13 : 3475-, 2009

5 Y. Liu, 7 : e33225-, 2012

6 D. L. Simpson, 109 : 274-, 2012

7 C. M. Crocco, 30 : 315-, 2013

8 E. Seyedjafari, 22 : 165-, 2011

9 I. Shabani, 17 : 2001

10 V. Beachley, 35 : 868-, 2000

11 W. T. Godbey, 961 : 10-, 2002

12 R. Cancedda, 22 : 81-, 2003

13 W. W. Thein-Wan, 5 : 3453-, 2009

14 K. Gauthaman, 13 : 3475-, 2009

15 M. J. Martin, 11 : 1-, 2005

16 A. Finne-Wistrand, 8 : 951-, 2008

17 H. Hajiali, 6 : 2133-, 2011

18 E. F. Irwin, 32 : 6912-, 2011

19 M. D. Pierschbacher, 309 : 30-, 1984

20 E. Ruoslahti, 12 : 697-, 1996

21 Y. C. Lim, 108 : 116-, 2011

22 S. Rungsiyanont, 22 : 1-, 2011

23 S.-C. Wu, 26 : 565-, 2011

24 E. Rosellini, 91 : 447-, 2009

25 E. Yasuda, 107 : 442-, 2009

26 C. Michon, 20 : 259-, 1997

27 Y. Tsao, 118 : 290-, 2005

28 C. Altamirano, 110 : 171-, 2004

29 C. Altamirano, 125 : 547-, 2006

30 J. Berrios, 66 : 2431-, 2011

31 T. Ryll, 44 : 184-, 1994

32 X. Gu, 58 : 643-, 1998

33 V. D. Shcherbukhin, 35 : 229-, 1999

34 V. D. Prajapati, 60 : 83-, 2013

35 M. Qingyuan, 33 : 1414-, 2012

36 F. T. Liu, 5 : 29-, 2005

37 D. Neri, 5 : 436-, 2005

38 N. L. Perillo, 185 : 1851-, 1997

39 M. C. Miller, 19 : 1034-, 2009

40 S. K. Bhatia, 24 : 315-, 2008

41 R. R. Ruggeri, 29 : 1039-, 2012

42 B. Follstad, "U.S. Patent No.2004/00810"

43 F. E. Noll, "U.S. Patent No. 2008/0220524 A1"

44 Heyun Wang, "Fabrication and Characterization of Electrospun Gelatin-Heparin Nanofibers as Vascular Tissue Engineering" 한국고분자학회 21 (21): 860-869, 2013

45 A. Choo, "Embryonic Stem Cell Therapy for Osteo-Degenerative Diseases, Methods in Biomolecular Biology, Vol. 690" Springer Science 175-182, 2011

46 Heyun Wang, "Electrospun Hemocompatible PU/ Gelatin-Heparin Nanofibrous Bilayer Scaffolds as Potential Artificial Blood Vessels" 한국고분자학회 20 (20): 347-350, 2012

47 고종현, "Characterization of Cross-linked Gelatin Nanofibers through Electrospinning" 한국고분자학회 18 (18): 137-143, 2010