알긴을 함유한 혼합 겔 비드 내에서 동물 세포의 고정화

이진호; 박종우; 이해방

Polymer(Korea), Vol.15, No.4, 474-480, August, 1991

Export Citation

알긴을 함유한 혼합 겔 비드 내에서 동물 세포의 고정화

Immobilization of Animal Cells in the Gel Beads of Alginate-Functional Polymer Mixture

이진호, 박종우, 이해방

초록

알긴과 여러가지 관능기를 갖는 고분자들의 혼합 겔 비드 채 에 동물세포인 CHO 세포와 BHK 세포를 고정화하여 이들 세포가 비드 내에서 살아있는 정도와 중식 경향을 고찰하였다. 혼합 겔 비드 제조를 위해 사용한 고분자들로서는 -OH기를 갖는 폴리비닐알코올, -COOH기를 갖는 폴리아크릴산,-NH₂기를 갖는 폴리아크릴 아미드 등이었다. 혼합 겔 비드 내에 고정화된 세포들은 37℃, 5% CO₂ 개스 분위기하에서 페트리 디쉬 내에서 배양되었다. 일정기간 배양후 비드 내에 살아있는 세포들의 밀도는 트리판블루 염색법과 혜모사이토메터를 사용하여 산출되었다. 비드 내부의 구조와 비드 내에서 자라고 있는 세포들은 SEM과 현미경에 의해 관찰되었다. 혼합 겔 비드들 중에서는 알긴과 폴리비닐알코올로 제조된 겔 비드가 세포 생존율과 중식성에 있어 다른 것들보다 우수한 것으로 판명되었다.

Animal cells, Chinese Hamster Ovary (CHO) and Baby Hamster Kidney (BHK) cells, were immobilized In the gel beads of alginate-functiollal Polymer mixture to investigate the effect of the mixtures of the alginate and the polymer with different functional group on the viability and growth of the cells. The polymers used were poly(vinyl alcohol) (PVA, -OH functional group), poly(acrylic acid) (PAA, -COOH functional group). and poly(acryl amide) (PAAM, -NH₂ functional group). The cells immobilized in the beads were cultured in Petri dishes at 37℃ under 5% CO₂ atmosphere. The viable cell densities In the beads after given time culture were measured by the trepan blue dye exclusion method using a haemocytometer. Morphologies of the beads and the cells growing in the beads were examined with a scanning electron microscope (SEM) and an inverted microscope. It was observed thats the beads prepared by the alginate-poly(vinyl alcohol) mixture are effective for the immobilization of both CHO and BHK cells in terms of cell viability and density in the beads after culturing in Petri dishes.

[References]

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[1] Nilsson K, Methods Enzymology, 135, 387 (1987)

[2] Tamponnet C, Boisseau S, Lirsac PN, Barbotin JN, Poujeol C, Lievremont M, Simonneau M, Appl. Microbiol. Biotechnol., 33, 442 (1990)

[3] Shirai Y, Hashimoto K, Yamaji H, Tokashiki M, Appl. Microbiol. Biotechnol., 26, 495 (1987)

[4] Lim F, U.S. Patent, 4,409,331 (1983)

[5] Nigam SC, Wang HY, PCT Int. Appl., WO 89/01034 (1989)

[6] Bucke C, Methods Enzymology, 135, 175 (1987)

[7] Lee JH, Park JW, Lee HB, Biomaterials, in press (1991)

[8] Lee JH, Khang GS, Park KH, Lee HB, Andrade JD, J. KOSOMBE, 10, 195 (1989)

[9] Jakoby WB, Pastan IH, "Methods in Enzymology. Vol. LVIII, Cell Culture," Academic Press, New York (1979)

[10] Shirai Y, Sasaki R, Hashimoto K, Kawahara H, Hitomi K, Chiba H, Appl. Microbiol. Biotechnol., 29, 544 (1988)

[11] Lee HY, Kang JK, HWAHAK KONGHAK, 27, 286 (1989)

[12] Goto M, Akai K, Murakami A, Hashimoto C, Tsuda E, Ueda M, Kawanishi G, Takahashi N, Ishimoto A, Chiba H, Sasaki R, Bio-Technol., 6, 67 (1988)

[13] Ryll T, Lnage ML, Jager V, Wagner R, J. Biotechnol., 14, 377 (1990)

[14] Croughan MS, Wang DIC, Biotechnol. Bioeng., 36, 316 (1990)

[15] Smiley AL, Hu WS, Wnag DIC, Biotechnol. Bioeng., 33, 1182 (1989)

[16] Kohn R, Pure Appl. Chem., 42, 371 (1975)