Ovine Fetal Adnexa-Derived Mesenchymal Stem Cells exhibit Stemness and Pluripotency with Distinction in Growth Kinetics and Clonogenic Potential

Fetal adnexa derived Mesenchymal Stem Cells (MSC) offer all in one solution in therapeutics. The present study was designed to isolate, cultivate, characterize and to evaluate the growth kinetics of ovine fetal adnexa-derived Mesenchymal Stem Cells (MSCs). The gravid uteri of ewes were collected from a local abattoir. The four cell lines viz. Wharton’s Jelly (oWJ), Cord Blood (oCB), Amniotic Fluid (oAF), and Amniotic Sac (oAS) were isolated and expanded in vitro and characterized for surface markers (CD73, CD90 & CD105) and pluripotency markers (SOX2 & OCT4) at 3rd passage. The growth kinetics of MSCs was investigated at 3rd and 5th passages. Similarly, the colony forming unit (CFU) Assay was performed at 3rd passage. The fetal adnexa derived Ovine MSCs showed the amplification of CD73, CD90 & CD105, whereas, negative marker CD34 was not amplified. Similarly, the MSCs also qualified for the amplification of Pluripotency markers (OCT4, SOX2). The MSCs in culture showed a typical growth curve with initial lag phase, an exponential phase, a plateau phase and a decline phase. The proliferation potential of MSCs was higher at P5 than P3. The overall growth rate was significantly (p < 0.05) highest in oAF, followed by oCB, oWJ and oAS. The population doubling time (PDT) was significantly (p < 0.05) highest in oAS (87.28 ± 3.24 hr) and least in oAF (39.75 ± 1.09 hr). The MSCs showed a significant (p < 0.05) difference in their clonogenic potential. The colony number was highest in oAF (53.67 ± 4.06) and least in oAS (22.0 ± 2.08). The study reveals that in terms of growth kinetics and clonogenic potential, oAF-MSCs were superior which outperformed other MSCs indicating that oAF derived MSCs could be utilized for regenerative medicine.