title: Expression and secretory profile of buffalo fetal fibroblasts and Wharton's jelly feeder layers.
abstract: The present study examined the comparative expression and secretory profile of vital signaling molecules in buffalo fetal fibroblasts (BFF) and Wharton's jelly (BWJ) feeder layers at different passages. Both feeder layers were expanded up to 8th passage. Signaling molecules viz. bone morphogenetic protein 4 (BMP4), fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF) and transforming growth factor beta 1 (TGFB1) and pluripotency-associated transcriptional factors (POU5F1, SOX2, NANOG, KLF4, MYC and FOXD3) were immunolocalized in the both feeder types. A clear variation in the expression pattern of key signaling molecules with passaging was registered in both feeders compared to primary culture (0 passage). The conditioned media (CM) was collected from different passages (2, 4, 6, 8) of both the feeder layers and was quantified using enzyme-linked immunosorbent assay (ELISA). Concomitant to expression profile, protein quantification also revealed differences in the concentration of signaling molecules at different time points. Conjointly, expression and secretory profile revealed that 2nd passage of BFF and 6th passage of BWJ exhibit optimal levels of key signaling molecules thus may be selected as best passages for embryonic stem cells (ESCs) propagation. Further, the effect of mitomycin-C (MMC) treatment on the expression profile of signaling molecules in the selected passages of BFF and BWJ revealed that MMC modulates the expression profile of these molecules. In conclusion, the results indicate that feeder layers vary in expression and secretory pattern of vital signaling molecules with passaging. Based on these findings, the appropriate feeder passages may be selected for the quality propagation of buffalo ESCs.
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Mehtab S
authors.initials: MS
authors.lastname: Parmar
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Smruti Ranjan
authors.initials: SR
authors.lastname: Mishra
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Anjali
authors.initials: A
authors.lastname: Somal
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Sriti
authors.initials: S
authors.lastname: Pandey
authors.affiliation: Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: G Sai
authors.initials: GS
authors.lastname: Kumar
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Mihir
authors.initials: M
authors.lastname: Sarkar
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India.
authors.forename: Vikash
authors.initials: V
authors.lastname: Chandra
authors.affiliation: Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, UP, India. Electronic address: gts553@gmail.com.
authors.forename: G Taru
authors.initials: GT
authors.lastname: Sharma
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D002020:Buffaloes; D018929:Cell Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D061252:Feeder Cells / Q000378:metabolism*; D005333:Fetus / Q000166:cytology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D016685:Mitomycin / Q000494:pharmacology; D059467:Transcriptome
chemical_list: D000477:Alkylating Agents; D016685:Mitomycin
keywords: Buffalo; Conditioned media; Feeder layers; Mitomycin; Signaling molecules