Leydig Cells Encapsulation with Alginate-Chitosan: Optimization of Microcapsule Formation


This research aimed to optimize the formation of microcapsules from alginate and chitosan for Leydig cells encapsulation. Alginate was used as the first coating agent while chitosan was the second layer. Various concentrations of alginate and CaCl2 were applied utilizing the extrusion method and the best concentration was determined based on their formation time, shape and diameter of microcapsules. Alginate microcapsule was applied with chitosan in various con- centrations. The best chitosan concentration was selected based on its mechanical stability. The results showed that the minimum concentration of alginate was 1.5% (w/v) with viscosity of 33.8 cPs, resulted to spherical microcapsules with diameters of 230 - 270 μm. The optimum concentration of chitosan as the second coating agent was 0.5% (w/v), resulted to spherical microcapsules with mechanical stability of 4 hours. Leydig cells were trapped inside the microcapsule with a density that is proportional to the concentration of cells used in the encapsulation.

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I. Batubara, D. Rahayu, K. Mohamad and W. Prasetyaningtyas, "Leydig Cells Encapsulation with Alginate-Chitosan: Optimization of Microcapsule Formation," Journal of Encapsulation and Adsorption Sciences, Vol. 2 No. 2, 2012, pp. 15-20. doi: 10.4236/jeas.2012.22003.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] M. L. Torre, M. Faustini, K. M. E. Attilio and D. Vigo, “Cell Encapsulation in Mammal Reproduction,” Recent Patents on Drug Delivery & Formulation, Vol. 1, No. 1, 2007, pp. 81-85. doi:10.2174/187221107779814078
[2] T. M. S. Chang, “Semipermeable Microcapsules,” Science, Vol. 146, No. 3643, 1964, pp. 524-525. doi:10.1126/science.146.3643.524
[3] M. Machluf, A. Orsola and A. Atala, “Controlled Release of Therapeutic Agents: Slow Delivery and Cell Encapsulation,” World Journal of Urology, Vol. 18, No. 1, 2000, pp. 80-83. doi:10.1007/s003450050014
[4] M. Machluf, A. Orsola, S. Boorjian, R. Kershen and A. Atala, “Microencapsultaion of Leydig Cells: A System for Testosterone Supplementation,” General Endocrinology, Vol. 144, No. 11, 2003, pp. 4975-4979. doi:10.1210/en.2003-0411
[5] B. Baxter, T. Eng and J. Zechlinski, “Microcapsulation of Cells,” BME 400 University of Wisconsin-Madison Final Report, Dec 7, 2005. pp 15-16. http://homepages.cae.wisc.edu/~bme200/microencapsulation_fall05/reports/BME_400_Microencapsulation_Final_Report.pdf
[6] A. C. Friedli and I. R. Schlager, “Demonstrating Encapsulation and Release: A New Take on Alginate Complexation and the Nylon Rope Trick,” Journal of Chemical Education, Vol. 82, No. 7, 2005, pp. 1017-1020. doi:10.1021/ed082p1017
[7] L. Capretto, S. Mazzitelli, G. Luca and C. Nastruzzi, “Preparation and Characterization of Polysaccharidic Microbeads by a Microfluidic Technique: Application to the Encapsulation of Sertoli Cells,” Acta Biomaterialia, Vol. 6, No. 2, 2010, pp. 429-435. doi:10.1016/j.actbio.2009.08.023
[8] C. M. Silva, A. J. Riberio, M. Figueiredo, D. Ferreira and F. Veiga, “Microencapsulation of Hemoglobin in Chitosan-Coated Alginate Microspheres Prepared by Emulsification/Internal Gelation,” The AAPS Journal, Vol. 7, No. 4, 2006, pp. E903-E913. doi:10.1208/aapsj070488
[9] J.-H. Zhu, X.-W. Wang, S. Ng, C.-H. Quek, H.-T. Ho, X.-J. Lao and H. Yu, “Encapsulating Live Cells with WaterSoluble Chitosan in Physiological Conditions,” Journal of Biotechnology, Vol. 117, No. 4, 2005, pp. 355-365. doi:10.1016/j.jbiotec.2005.03.011
[10] H. Chemes, S. Cigorraga, C. Begadá, H. Schteingart, R. Rey and E. Pellizzari, “Isolation of Human Leydig Cell Mesenchymal Precursors from Patient with the Androgen Insensitivity Syndrome: Testosterone Production and Response to Human Chorionic Gonadotropin Stimulation in Culture,” Biology of Reproduction, Vol. 46, No. 5, 1992, pp. 793-801. doi:10.1095/biolreprod46.5.793
[11] V. T. Nguyen, S. Kure-Bayashi, H. Harayama, T. Nagai and M. Miyake, “Stage Spesific Effects of the Osmolality of a Culture Medium on the Development of Parthenogenetic Diploids in Pig” Theriogenology, Vol. 59, No. 3, 2003, pp. 719-734. doi:10.1016/S0093-691X(02)01085-3
[12] G. Murtaza, M. Ahamd, N. Akhtar and F. Rasool, “A Comparative Study of Various Microencapsulation Techniques: Effect of Polymer Viscosity on Microcapsule Characteristics,” Pakistan Journal of Pharmaceutical Science, Vol. 22, No. 3, 2009, pp. 291-300.
[13] M. F. A. Goosen, G. M. O’Shea and M. F. Sun, “Microencapsulation of Living Tissue and Cells,” US Patent No. 4,806,355, 1998.
[14] http://www.google.com/patents?id=KARLAAAAEBAJ& pg=PA6&dq=microcapsules+of+living+tissue&hl=en&sa=X&ei=ZozNT6CiFI_imAWKv7G4Aw&ved=0CDQQ6 AEwAA#v=onepage&q=microcapsules%20of%20living %20tissue&f=false
[15] I. Stuiver, “Microencapsulation of Islet for the Treatment of Type 1 Diabetes,” Workshop Encapsulation and Immunoprotective Strategies of Islet Cells Proceeding, Washington DC, 6-7 December 2001, p. 18. H. Uludag, P. De Vos and P. A. Tresco, “Technology of Mammalian Cell Encapsulation,” Advanced Drug Delivery Reviews, Vol. 42, No. 1-2, 2000, pp. 29-64. doi:10.1016/S0169-409X(00)00053-3

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