Method for culturing differentiation-promoting and -sustaining spheroid form of tonsil-derived stem cells

A method for producing a spheroid form of parathyroid cells from tonsil-derived stem cells, the method comprising:

• (a) transferring a suspension of tonsil-derived stem cells into a hemispherical microwell that is formed in such a way that at least one liquid polymer is hardened in a form of hemispherical microwell, wherein the liquid polymer is selected from among polydimethylsiloxane, silicon polymer, polybutadiene, polyisobutylene and polyurethane and forms a meniscus due to surface tension;
• (b) producing a spheroid form of tonsil-derived stem cells by culturing the tonsil-derived stem cells prepared by the step (a) in the hemispherical microwell;
• and
• (c) adding activin A and sonic hedgehog into the spheroid form of tonsil-derived stem cells of the step (b), thereby differentiating the stem cells into parathyroid cells.

A method for producing parathyroid hormone from tonsil-derived stem cells, the method comprising:

• (a) transferring a suspension of tonsil-derived stem cells into a hemispherical microwell that is formed in such a way that at least one liquid polymer is hardened in a form of hemispherical microwell, wherein the liquid polymer is selected from among polydimethylsiloxane, silicon polymer, polybutadiene, polyisobutylene and polyurethane and forms a meniscus due to surface tension;
• (b) producing a spheroid form of tonsil-derived stem cells by culturing the tonsil-derived stem cells prepared by the step (a) in the hemispherical microwell;
• (c) adding activin A and sonic hedgehog into the spheroid form of tonsil-derived stem cells of the step (b), thereby differentiating the stem cells into parathyroid cells;
• and
• (d) separating parathyroid hormone from culture medium or cell lysis solution obtained from the step (c).