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




Abstract

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

References

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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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