Phenotype and multipotency of rabbit (Oryctolagus cuniculus) amniotic stem cells.
Stem cells are capable of unlimited self-renewal and are able to remain undifferentiated for extended periods of time prior to their differentiation into specific cell lineages. Because of the issues (ethical and religious) involved in the use of embryonic stem cells and the limited plasticity of adult stem cells, an alternative cell source could be foetal stem cells derived from extra-embryonic tissue, which are highly proliferative, grow in vitro and possess interesting immunogenic characteristics. As a result, the amniotic membrane of several species has been studied as an important new source of stem cells.



Here, we cultured and characterized mesenchymal progenitor cells derived from the rabbit amniotic membrane, and investigated their differentiation potential. In total, amniotic membranes were collected from eight rabbit foetuses and were isolated by the explant technique. The obtained cells were cultured in DMEM-HIGH glucose and incubated at 37 °C in a humidified atmosphere with 5% CO2.



The cells adhered to the culture plates and showed a high proliferative capacity with fibroblast-like morphologies. The cells showed a positive response for markers for the cytoskeleton, mesenchymal stem cells and proliferation, pluripotency and haematopoietic precursor stem cells. However, the cells were negative for CD45, a marker of haematopoietic cells. Furthermore, the cells had the capacity to be induced to differentiate into osteogenic, adipogenic and chondrogenic lineages. In addition, when the cells were injected into nude mice, we did not observe the formation of tumours.



In summary, our results demonstrate that multipotent mesenchymal stem cells can be obtained from the rabbit amniotic membrane for possible use in future cell therapy applications.
Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, SP, Brazil. jehborghesi@hotmail.com.|Orlando Marques de Paiva, 87, Cidade Universitária, Sao Paulo, SP, 05508-270, Brazil. jehborghesi@hotmail.com.
Jéssica
J
Borghesi
Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, SP, Brazil.
Lara Carolina
LC
Mario
NUCEL (Cell and Molecular Therapy Center) and NETCEM (Center for Studies in Cell and Molecular Therapy), School of Medicine-Chemistry Institute, Biochemistry Department, Sao Paulo University, Sao Paulo, SP, Brazil.
Ana Claudia Oliveira
AC
Carreira
Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, SP, Brazil.
Maria Angélica
MA
Miglino
Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, SP, Brazil.
Phelipe Oliveira
PO
Favaron
D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000650:Amnion / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010641:Phenotype; D061251:Primary Cell Culture; D011817:Rabbits
D003470:Culture Media; D005947:Glucose
Amniotic membrane; Mesenchymal stem cells; Proliferative capacity; Rabbit