Effect of Different Factors on Proliferation of Antler Cells, Cultured In Vitro

Antlers as a potential model for bone growth and development have become an object of rising interest. To elucidate processes explaining how antler growth is regulated, in vitro cultures have been established. However, until now, there has been no standard method to cultivate antler cells and in vitro results are often opposite to those reported in vivo. In addition, many factors which are often not taken into account under in vitro conditions may play an important role in the development of antler cells. In this study we investigated the effects of the antler growth stage, the male individuality, passaged versus primary cultures and the effect of foetal calf serum concentrations on proliferative potential of mixed antler cell cultures in vitro, derived from regenerating antlers of red deer males (Cervus elaphus). The proliferation potential of antler cells was measured by incorporation of 3H thymidine. Our results demonstrate that there is no significant effect of the antler growth stage, whereas male individuality and all other examined factors significantly affected antler cell proliferation. Furthermore, our results suggest that primary cultures may better represent in vivo conditions and processes occurring in regenerating antlers. In conclusion, before all main factors affecting antler cell proliferation in vitro will be satisfactorily investigated, results of in vitro studies focused on hormonal regulation of antler growth should be taken with extreme caution

Hormonal Aspects of Antler Growth Regulation

Hormonal aspects of antler growth regulation Erika Kužmová Abstract Deer antlers are the only mammalian organ that completely regenerates and therefore they became an object of rising interest as a potential model for bone growth and development. In recent years, it has been confirmed that annual regeneration of the antler is initiated from the stem cell niche localised in the pedicle periosteum. Antlers grow to the length at the tip. Only a little is known about endocrine stimulation of antler growth and some discrepancy has arisen between in vivo and in vitro studies over the decades. As the secondary sexual character, the antler cycle timing and growth are linked to seasonal levels of testosterone. Since the levels are at their minimum during the antler growth phase, according to many mainly in vitro studies, insulin-like growth factor-1 (IGF-1) tends to be accepted as the "antler stimulating hormone". Since the conclusion about the role of IGF-1 was contradictory to previous opinions and also in contrast with our own experience, we aimed to verify the role of IGF-1 in vitro. Our ex- periments were based on existing in vivo studies demonstrating the importance of testosterone, even in its low levels, and on the hypothesis that testosterone should be the "antler stimulating hormone". We performed in vitro..

Hormonal Aspects of Antler Growth Regulation

Hormonal aspects of antler growth regulation Erika Kužmová Abstract Deer antlers are the only mammalian organ that completely regenerates and therefore they became an object of rising interest as a potential model for bone growth and development. In recent years, it has been confirmed that annual regeneration of the antler is initiated from the stem cell niche localised in the pedicle periosteum. Antlers grow to the length at the tip. Only a little is known about endocrine stimulation of antler growth and some discrepancy has arisen between in vivo and in vitro studies over the decades. As the secondary sexual character, the antler cycle timing and growth are linked to seasonal levels of testosterone. Since the levels are at their minimum during the antler growth phase, according to many mainly in vitro studies, insulin-like growth factor-1 (IGF-1) tends to be accepted as the "antler stimulating hormone". Since the conclusion about the role of IGF-1 was contradictory to previous opinions and also in contrast with our own experience, we aimed to verify the role of IGF-1 in vitro. Our ex- periments were based on existing in vivo studies demonstrating the importance of testosterone, even in its low levels, and on the hypothesis that testosterone should be the "antler stimulating hormone". We performed in vitro..

Hormonální aspekty regulace parožního růstu

Hormonal aspects of antler growth regulation Erika Kužmová Abstract Deer antlers are the only mammalian organ that completely regenerates and therefore they became an object of rising interest as a potential model for bone growth and development. In recent years, it has been confirmed that annual regeneration of the antler is initiated from the stem cell niche localised in the pedicle periosteum. Antlers grow to the length at the tip. Only a little is known about endocrine stimulation of antler growth and some discrepancy has arisen between in vivo and in vitro studies over the decades. As the secondary sexual character, the antler cycle timing and growth are linked to seasonal levels of testosterone. Since the levels are at their minimum during the antler growth phase, according to many mainly in vitro studies, insulin-like growth factor-1 (IGF-1) tends to be accepted as the "antler stimulating hormone". Since the conclusion about the role of IGF-1 was contradictory to previous opinions and also in contrast with our own experience, we aimed to verify the role of IGF-1 in vitro. Our ex- periments were based on existing in vivo studies demonstrating the importance of testosterone, even in its low levels, and on the hypothesis that testosterone should be the "antler stimulating hormone". We performed in vitro...Hormonálne aspekty regulácie parožného rastu Erika Kužmová Abstract Parohy jeleňov sú jediným kompletne sa regenerujúcim orgánom u cicavcov a záujem vedcov o ich využitie ako modelu rastu a vývoja kostí stúpa. V posledných rokoch sa ukázalo, že regenerácia parohov je iniciovaná z kmeňových buniek lokalizovaných v okostici pučnice. Následný rast parohu do dĺžky však prebieha v rastovom vrcholčeku. Len málo sa vie o endokrinnej stimulácii rastu parohu a už dlhé roky existuje nesúlad medzi in vivo a in vitro štúdiami. Ako druhotný sexuálny znak sú parohy úzko späté so sezónnymi hladinami cirkulujúceho testosterónu. Keďže sú jeho hladiny najnižšie práve v čase rastu parohov a mnohé in vitro štúdie poukazujú na stimulačný efekt inzulínu podobného rastového faktoru (IGF-1), viacerí odborníci sa prikláňajú k názoru že IGF-1 je "hormón stimulujúci rast parohov". Tento záver je ale v rozpore s výsledkami in vivo štúdií, ktoré ukazujú nevyhnutnosť testosterónu pre rast parohov aj v jeho nízkych koncentráciách, a taktiež s predchádzajúcim názorom, že testosterón by mal byť "hormón stimulujúci rast parohov". Zamerali sme sa teda na overenie účinkov IGF-1 na parožné bunky. Uskutočnili sme sériu in vitro experimentov na parožných bunkách izolovaných z viacerých štádií rastových vrcholčekov parohov jeleňa európskeho...Department of EcologyKatedra ekologieFaculty of SciencePřírodovědecká fakult

Hormonal Aspects of Antler Growth Regulation

Hormonal aspects of antler growth regulation Erika Kužmová Abstract Deer antlers are the only mammalian organ that completely regenerates and therefore they became an object of rising interest as a potential model for bone growth and development. In recent years, it has been confirmed that annual regeneration of the antler is initiated from the stem cell niche localised in the pedicle periosteum. Antlers grow to the length at the tip. Only a little is known about endocrine stimulation of antler growth and some discrepancy has arisen between in vivo and in vitro studies over the decades. As the secondary sexual character, the antler cycle timing and growth are linked to seasonal levels of testosterone. Since the levels are at their minimum during the antler growth phase, according to many mainly in vitro studies, insulin-like growth factor-1 (IGF-1) tends to be accepted as the "antler stimulating hormone". Since the conclusion about the role of IGF-1 was contradictory to previous opinions and also in contrast with our own experience, we aimed to verify the role of IGF-1 in vitro. Our ex- periments were based on existing in vivo studies demonstrating the importance of testosterone, even in its low levels, and on the hypothesis that testosterone should be the "antler stimulating hormone". We performed in vitro..

Effect of the FCS on the antler cell proliferation.

<p>Incorporation of ³H thymidine in antler cells (least square means ± S.E.) according to FCS percentage. All other factors were statistically eliminated.</p

Effect of the individual males on the antler cell proliferation.

<p>Incorporation of ³H thymidine in antler cells (least square means ± S.E.) according to the individual males (A, B, C). All other factors were statistically eliminated.</p

Effect of the passage on the antler cell proliferation.

<p>Incorporation of ³H thymidine in antler cells (least square means ± S.E.) according to the passage (primary culture, passaged culture – 2^nd passage). All other factors were statistically eliminated.</p

Tissue sampling.

<p>Example of tissue sampling from anesthetized animals using a sterile trephine punch.</p