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CELL CONCENTRATION AND FENOTIPIC DIFFERENCES BETWEEN MESENCHYMAL CELLS IN HUMAN, SHEEP AND RABBIT.



Abstract

Introduction. Progenitor cells with osteochondrogenic potential have been identified within adipose tissue. These cells present diversity of characteristics that can be explained by differences in tissue origin, isolation methods and culture conditions. Mesenchymal stem cells (MSC) have been isolated from many tissues. MSC have been shown to exhibit tissue protective and regenerative properties.

Methods. Hoffa’s fat samples were obtained from four patients (mean age 44 years), five rabbits (New Zeland aged 12 weeks) and five sheeps (Rasa aragonesa aged 22 weeks). Cells were obtained by means of enzimatic and mechanical digestion. The suspension was centrifuged and washed twice with phosphate buffered saline. The resultant pellet was resuspended and plated in culture medium (37°C, CO2 5%). Cellular markers were studied with specific monoclonal antibodies (CD13, CD44, CD49d, CD90, CD105, CD117).

Results:

  • Human cells: CD13+ (94–99%), CD44+ (87–99), CD49d (14–70%), CD90+ (92–99%), CD105+ (90–97%), CD 117-BD+ (2–22%).

  • Sheep cells presented CD13+ (32–70%), CD34-, CD36, CD44+ (90–96%), CD49d (40–80%), CD54+ (50–80%), CD90+ (90–97%), CD105+ (10–25%). CD117-BD+ (48–76%).

  • Rabbits cells: CD13+ (14–78%), CD44+ (10–80%), CD49d (2–9%), CD90+ (27–92%), CD105+ (2–24%), CD 117-BD+ (15–57%). Human cells number/mL did not show significant differences between patients, or between P0 0 (14 culture days) (average mean: 525000 ± 298956) and P5 (525000), nevertheless the average mean decreased from P5 to P6 (130.000) until P8 (111 culture days) (85.000). Rabbits cells number/mL did not show significant differences between P0 (673000 ± 379697) and P1 (596000 ± 488740) and decreased in P2 (299500 ± 159161) without any significant change in P8. Ovine cells number/mL average mean in P0 was 1.370.600 (± 802758), this decreased in P1 (420000 ± 95197) however, showed no significant changes in P8 (291875 ± 86394).

Conclusions: MSC from human, rabbits and sheeps present differences in cellular concentration and markers.

Correspondence should be addressed to EORS Secretariat Mag. Gerlinde M. Jahn, c/o Vienna Medical Academy, Alserstrasse 4, 1090 Vienna, Austria. Fax: +43-1-4078274. Email: eors@medacad.org