Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Background: In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab‐to‐lab variability jeopardizes the much‐needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources. Methods: The most commonly applied methods for NP cell extraction, expansion, and re‐differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re‐differentiation media and techniques were also investigated. Results: Recommended protocols are provided for extraction, expansion, and re‐differentiation of NP cells from common species utilized for NP cell culture. Conclusions: This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species‐specific pronase usage, 60–100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross‐lab comparisons on NP cells worldwide

Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Background In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab-to-lab variability jeopardizes the much-needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources. Methods The most commonly applied methods for NP cell extraction, expansion, and re-differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re-differentiation media and techniques were also investigated. Results Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture. Conclusions This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species-specific pronase usage, 60–100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross-lab comparisons on NP cells worldwide

Stakeholder Involvement in Outbreak Management: To Fear or Not to Fear?

Infectious diseases remain a threat to public health, requiring the coordinated action of many stakeholders. Little has been written about stakeholder participation and approaches to sharing information, in dynamic contexts and under time pressure as is the case for infectious disease outbreaks. Communicable-disease specialists fear that delays in implementing control measures may occur if stakeholders are not included in the outbreak-management process. Two case studies described in this article show how the needs of stakeholders may vary with time and that early sharing of information takes priority over shared decision-making. The stakeholders itemized their needs and potential contributions in order to arrive at the collective interest of outbreak management. For this, the results suggest the potential for improvement through development of “network governance” including the effective sharing of information in large networks with varying needs. Outbreaks in which conflicting perceptions may occur among the stakeholders require particular attention

Ebola Preparedness in the Netherlands: The Need for Coordination Between the Public Health and the Curative Sector.

During the Ebola outbreak in West Africa in 2014-2015, close cooperation between the curative sector and the public health sector in the Netherlands was necessary for timely identification, referral, and investigation of patients with suspected Ebola virus disease (EVD)

Knowledge, perceptions and media use of the Dutch general public and healthcare workers regarding Ebola, 2014.

The Ebola outbreak in West-Africa triggered risk communication activities to promote adequate preventive behaviour in the Netherlands. Our study investigated the level of knowledge, perceptions, and media use regarding Ebola