Research Article
Volume 3 Issue 9 - 2020
Isolation of Theca Cells from Goat Ovarian Follicles and Expression of CYP19 Gene in the Isolated Cells
Anjana A1, Raji K2*, Bhuvana Plakkot3 and Aravindhakshan TV4
1Former MSc Student, School of Applied Animal Production and Biotechnology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India
2Associate Professor, Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India
3Former MVSc Scholar, Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India
4Director, School of Applied Animal Production and Biotechnology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India
*Corresponding Author: Raji K, Associate Professor, Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India.
Received: May 21, 2020; Published: August 31, 2020


Ovarian Theca Cells (TCs) isolation is quite difficult as they are attached to connective tissue and usually isolated TCs have contamination with Granulosa cells (GCs). Contamination of TCs with GCs can be detected by the marker genes which are expressed only in GCs. Cytochrome P450 aromatase (CYP 19) is a marker for GCs which is responsible for the formation of enzyme aromatase for the synthesis of oestradiol from androgen. This study was undertaken to isolate TCs from caprine ovarian follicles by two different methods and to assess the efficacy of these methods to isolate theca cells with minimum contamination by GCs. In the study, follicles measuring three to six-millimetre diameter were used for isolation of TCs. Theca cells were isolated by two different methods - Trypsin and Collagenase methods. In the first method [1], trypsin with hyaluronidase was used and in the second method [2] different concentrations and combinations of collagenase and hyaluronidase enzymes were used. The isolated TCs were visualised under the inverted microscope and samples from both the methods were subjected to real-time qPCR for the presence of GCs specific marker Cytochrome P450 aromatase (CYP 19). Cytochrome P450 17 α-hydroxylase (CYP 17), marker for TC, gene expression was also studied to estimate the yield of TC by two methods. From this study, it was confirmed that collagenase was the better method for TCs isolation.

Keywords: Caprine Ovarian Follicles; Theca Cells; CYP 17; CYP 19; qRT-PCR


  1. Stoklosowa S., et al. “Some morphological and functional characteristics of cells of the porcine thecal and granulosa cells”. Biology of Reproduction 19 (1978): 712-719.
  2. Kataoka N., et al. “Preparation of highly purified porcine theca cells”. Journal of Reproduction and Fertility 102 (1994): 73-79.
  3. Zhou W., et al. “Effect of bisphenol A on steroid hormone production in rat ovarian theca-interstitial and granulosa cells”. Molecular and Cellular Endocrinology 283 (2008): 12-18.
  4. McNatty KP., et al. “Control of early ovarian follicular development”. Journal of Reproduction and Fertility Supplement 54 (1999): 3-16.
  5. Edson MA., et al. “The mammalian ovary from genesis to revelation”. Endocrine Reviews 30 (2009): 624-712.
  6. Araujo VR., et al. “In vitro culture of bovine preantral follicles: a review”. Reproductive Biology and Endocrinology 12 (2014): 78.
  7. Hatzirodos N., et al. “Transcriptome comparisons identify new cell Marker for theca Interna and Granulosa cells from small and large antral ovarian follicles”. PLoS ONE 3 (2015): 1-13.
  8. Livak KJ and Schmittgen TD. “Analysis of relative gene expression data using real-time quantitative PCR and the 22DDCT method”. Methods 25 (2001): 402-408.
  9. Sangha GK., et al. “Morphological changes in cultured granulosa cells from different sized follicles of goat ovary”. Indian Journal of Animal Reproduction1 (2012): 1-5.
  10. Li KB and Hearn TW. “Isolation of theca cells: an assessment of purity and steroidogenic potential”. The Journal of Biochemical and Biophysical Methods 45 (2000): 169-181.
  11. Dissen GA., et al. “Direct Effects of Nerve Growth Factor on Thecal Cells from Antral Ovarian Follicles”. Endocrinology 12 (2000): 4736-4750.
  12. Liu X., et al. “Paracrine regulation of steroidogenesis in theca cells by granulosa cells derived from mouse preantral follicles”. BioMed Research International (2015): 925691.
  13. Erickson GF., et al. “The ovarian androgen producing cells: a review of structure/function relationships”. Endocrine Reviews (1985): 6371-6399.
  14. Kotsuji F., et al. “Bovine theca and granulosa cell interactions modulate their growth, morphology, and function”. Biology of Reproduction 43 (1990): 726-732.
  15. Hsueh AJ., et al. “Ovarian follicle atresia: a hormonally controlled apoptotic process”. Endocrine Reviews 15 (1994): 707-724.
  16. Tian Y., et al. “Isolation and identification of ovarian theca-interstitial cells and granulosa cells of immature female mice”. Cell Biology International 39 (2015): 584-590.
  17. Simpson ER. “Aromatase: biologic relevance of tissue-specific expression”. Seminars in Reproductive Medicine 1 (2004): 11-23.
  18. Skinner MK., et al. “Regulation of granulosa and theca cell transcriptomes Ovarian antral follicle development”. Molecular Reproduction and Development 9 (2008): 1457-1472.
Citation: Raji K., et al. “Isolation of Theca Cells from Goat Ovarian Follicles and Expression of CYP19 Gene in the Isolated Cells”. EC Clinical and Experimental Anatomy 3.9 (2020): 10-19.

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