title: Impact of Cryopreservation on Caprine Fetal Adnexa Derived Stem Cells and Its Evaluation for Growth Kinetics, Phenotypic Characterization, and Wound Healing Potential in Xenogenic Rat Model.
abstract: This study was conducted to know the impact of cryopreservation on caprine fetal adnexa derived mesenchymal stem cells (MSCs) on the basic stem cell characteristics. Gravid caprine uteri (2-3 months) were collected from local abattoir to derive (amniotic fluid [cAF], amniotic sac [cAS], Wharton's jelly [cWJ], and cord blood [cCB]) MSCs and expanded in vitro. Cells were cryopreserved at 3rd passage (P3) using 10% DMSO. Post-thaw viability and cellular properties were assessed. Cells were expanded to determine growth kinetics, tri-lineage differentiation, localization, and molecular expression of MSCs and pluripotency markers; thereafter, these cells were transplanted in the full-thickness (2 × 2cm2) rat skin wound to determine their wound healing potential. The post-thaw (pt) growth kinetics study suggested that cWJ MSCs expanded more rapidly with faster population doubling time (PDT) than that of other fetal adnexa MSCs. The relative mRNA expression of surface antigens (CD73, CD90, and CD 105) and pluripotency markers (Oct4, KLF, and cMyc) was higher in cWJ MSCs in comparison to cAS, cAF, and cCB MSCs post-thaw. The percent wound contraction on 7th day was more than 50% for all the MSC-treated groups (pre and post-thaw), against 39.55% in the control group. On day 28th, 99% and more wound contraction was observed in cAF, cAF-pt, cAS-pt, cWJ, cWJ-pt, and cCB, MSCs with better scores for epithelization, neovascularization, and collagen characteristics at a non-significant level. It is concluded that these MSCs could be successfully cryopreserved without altering their stemness and wound healing properties. J. Cell. Physiol. 232: 2186-2200, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Anjali
authors.initials: A
authors.lastname: Somal
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Irfan A
authors.initials: IA
authors.lastname: Bhat
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Indu
authors.initials: I
authors.lastname: B
authors.affiliation: Division of Surgery, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Anuj P
authors.initials: AP
authors.lastname: Singh
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Bibhudatta S K
authors.initials: BSK
authors.lastname: Panda
authors.affiliation: Division of Veterinary Pathology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Perumal A
authors.initials: PA
authors.lastname: Desingu
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Sriti
authors.initials: S
authors.lastname: Pandey
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Mukesh K
authors.initials: MK
authors.lastname: Bharti
authors.affiliation: Division of Surgery, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Amar
authors.initials: A
authors.lastname: Pal
authors.affiliation: Division of Veterinary Pathology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Guttula
authors.initials: G
authors.lastname: Saikumar
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Vikash
authors.initials: V
authors.lastname: Chandra
authors.affiliation: Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India.
authors.forename: Guttula Taru
authors.identifier: http://orcid.org/0000-0002-5052-9477
authors.initials: GT
authors.lastname: Sharma
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D036101:Cord Blood Stem Cell Transplantation; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004195:Disease Models, Animal; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D053686:Fetal Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000637:transplantation*; D006041:Goats; D064593:Heterografts; D007700:Kinetics; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism; D010641:Phenotype; D011247:Pregnancy; D017208:Rats, Wistar; D000072836:Surgical Wound / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D059631:Wharton Jelly / Q000166:cytology; D014945:Wound Healing
chemical_list: D015415:Biomarkers; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide