Growth of recombinant fibroblasts in alginate microcapsules

P. L. Chang, G. Hortelano, M. Tse, D. E. Awrey

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

To develop a novel strategy of nonautologous somatic gene therapy, we now demonstrate the feasibility of culturing genetically modified fibroblasts within an immunoprotective environment and the optimal conditions required for their continued survival in vitro. When mouse Ltk- fibroblasts transfected with the human growth hormone gene were enclosed within permselective microcapsules fabricated from alginate-polylysine-alginate, they continued to secrete human growth hormone at the same rates as the nonencapsulated cells. They also continued to proliferate in vitro for at least 1 month even though their viability gradually declined to about 50%. The viability can be improved by controlling for (a) temperature during encapsulation, (b) duration of treatment with polylysine, (c) duration of liquefying the core alginate with sodium citrate, and (d) cell density at the time of encapsulation. The best conditions leading to improved survival and maximum proliferation of cells within the microcapsules were obtained by encapsulating the cells at 4 to 10°C instead of room temperature, coating the microspheres with polylysine for 6 to 10 min instead of 20 min, liquefying the core alginate by treating with citrate for 20 min instead of 6 to 10 min, and using a concentration of 2 x 106 cells/mL of alginate for encapsulation. Under such conditions, normally adherent and genetically engineered mouse fibroblasts survived and proliferated optimally within the microcapsule environment. The encapsulated fibroblasts maintained their level of transgene expression while recombinant gene products such as human growth hormone could diffuse through the microcapsule membrane without impediment. The demonstration that genetically modified fibroblasts can survive and continue to deliver recombinant gene products from within these microcapsules and the optimization for their maximal viability and growth within microcapsules should increase the potential for success in using such microencapsulated recombinant cells for somatic gene therapy.

Original languageEnglish
Pages (from-to)925-933
Number of pages9
JournalBiotechnology and Bioengineering
Volume43
Issue number10
DOIs
Publication statusPublished - 1994
Externally publishedYes

Fingerprint

Alginate
Fibroblasts
Capsules
Hormones
Encapsulation
Human Growth Hormone
Growth
Gene therapy
Genes
Polylysine
Genetic Therapy
Temperature
Microspheres
Transgenes
Citric Acid
Demonstrations
Sodium
alginic acid
Membranes
Cell Count

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

Cite this

Growth of recombinant fibroblasts in alginate microcapsules. / Chang, P. L.; Hortelano, G.; Tse, M.; Awrey, D. E.

In: Biotechnology and Bioengineering, Vol. 43, No. 10, 1994, p. 925-933.

Research output: Contribution to journalArticle

Chang, P. L. ; Hortelano, G. ; Tse, M. ; Awrey, D. E. / Growth of recombinant fibroblasts in alginate microcapsules. In: Biotechnology and Bioengineering. 1994 ; Vol. 43, No. 10. pp. 925-933.
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