Mesenchymal stem cells (MSC) can be isolated from a variety of adult animal tissues, replicated in vitro and induced to differentiate into different cell lineages. In the dog many districts have been investigated for MSC but never the female genital tract. In present paper we simultaneously studied the tissue collected from three different sites: ovarian bursa adipose tissue, endometrium and abdominal adipose tissue, used as control. We considered 7 intact bitches with the following inclusion criteria: primipary/nulliparous, ranging in age from 1 to 3 years, clinically healthy, weighing 1030 kg. Samples of abdominal adipose tissue, uterine horn (middle portion) and ovarian bursa were collected in a sterile environment during surgical procedure for elective ovariohysterectomy. Isolation, expansion and differentiation were carried out according to Bunnell et al (2008). Total RNA was extracted and cDNA were synthesized from 2 µg RNA to analyze the expression of marker typical of MSC: CD44, CD29, CD90, CD13, CD133, CD73, CD105, Oct4, CD45, CD31, CD34, GAPDH. RT-PCR were performed with these cycling condition: annealing 35 cycles at 55°C for 30 sec; denaturation at 94°C for 30 sec; extension at 72°C for 30 sec. Osteocytes, chondrocytes and adipocytes have been isolated from endometrium and abdominal adipose tissue (4 bitches), from ovarian bursa adipose tissue only two lines were tested (5 bitches). All isolated cells were able to adhere to plastic support generating colony forming units, showed similar fibroblastic-like morphology and ability of subculture; MSC isolated from abdominal adipose tissue and endometrium showed plasticity and expressed specific surface antigens. Cells isolated from ovarian bursa adipose tissue did not express specific surface antigens and did not show plasticity. We reported for the first time isolation and characterization of MSC from canine endometrium, a tissue that after ovariohysterectomy is routinely discharged and the rate of success for MSC isolated from this tissue was 100%.
CULTURE OF CANINE MESENCHYMAL STEM CELLS FROM OVARIAN BURSA ADIPOSE TISSUE, ENDOMETRIUM AND ABDOMINAL ADIPOSE TISSUE: A COMPARATIVE STUDY / de Cesaris, V.; Grolli, S.; Bertocchi, M.; Conti, V.; Bresciani, C.; di Ianni, F.; Parmigiani, E.; Bigliardi, E.. - (2016), pp. 368-368.
CULTURE OF CANINE MESENCHYMAL STEM CELLS FROM OVARIAN BURSA ADIPOSE TISSUE, ENDOMETRIUM AND ABDOMINAL ADIPOSE TISSUE: A COMPARATIVE STUDY
de Cesaris V.
;Grolli S.;Bertocchi M.;Conti V.;Bresciani C.;di Ianni F.;Parmigiani E.;Bigliardi E.
2016-01-01
Abstract
Mesenchymal stem cells (MSC) can be isolated from a variety of adult animal tissues, replicated in vitro and induced to differentiate into different cell lineages. In the dog many districts have been investigated for MSC but never the female genital tract. In present paper we simultaneously studied the tissue collected from three different sites: ovarian bursa adipose tissue, endometrium and abdominal adipose tissue, used as control. We considered 7 intact bitches with the following inclusion criteria: primipary/nulliparous, ranging in age from 1 to 3 years, clinically healthy, weighing 1030 kg. Samples of abdominal adipose tissue, uterine horn (middle portion) and ovarian bursa were collected in a sterile environment during surgical procedure for elective ovariohysterectomy. Isolation, expansion and differentiation were carried out according to Bunnell et al (2008). Total RNA was extracted and cDNA were synthesized from 2 µg RNA to analyze the expression of marker typical of MSC: CD44, CD29, CD90, CD13, CD133, CD73, CD105, Oct4, CD45, CD31, CD34, GAPDH. RT-PCR were performed with these cycling condition: annealing 35 cycles at 55°C for 30 sec; denaturation at 94°C for 30 sec; extension at 72°C for 30 sec. Osteocytes, chondrocytes and adipocytes have been isolated from endometrium and abdominal adipose tissue (4 bitches), from ovarian bursa adipose tissue only two lines were tested (5 bitches). All isolated cells were able to adhere to plastic support generating colony forming units, showed similar fibroblastic-like morphology and ability of subculture; MSC isolated from abdominal adipose tissue and endometrium showed plasticity and expressed specific surface antigens. Cells isolated from ovarian bursa adipose tissue did not express specific surface antigens and did not show plasticity. We reported for the first time isolation and characterization of MSC from canine endometrium, a tissue that after ovariohysterectomy is routinely discharged and the rate of success for MSC isolated from this tissue was 100%.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
