Abstract
High efficacy of placental cells application necessitates their investigation. Preclinical studies require an improvement of the methods for obtaining, standardizing and storage of placental cells of experimental animals. Cells were isolated from rats and mice placentas by means of different enzymatic methods and the one of explants. Cells were cryopreserved with DMSO in DMEM using two-stage freezing. The number, morphological, cultural, metabolic features of cells were studied after isolation and storage. The maximum number of viable cells from the placentas of mice and rats was found to be obtained using the explant method or trypsin with ETDA. Cell cultures from mice and rats placentas after the third passage had stable morphofunctional characteristics. The viability of warmed rat placental cells according to dye exclusion was (92.3 ± 1.6)%, according to the adhesive test this was (81.3 ± 5.8)%. For mice placental cells, these values were (86.7 ± 3.7)% and (79.2 ± 8.1)%, correspondingly. The research results enabled the determining of effective biotechniques for obtaining the cryopreserved placental cells of rats and mice to perform preclinical studies of their biological effect in models of allo- and autotransplantations.
Highlights
A high efficacy of placental cells application necessitates their investigation
The findings demonstrated that the largest number of cells from the placentas of rats and mice can be obtained by explants and enzymatic disaggregation by trypsin with ETDA or collagenase (Fig. 1A)
The maximum percentage of viable cells was obtained by the explant culture, the minimum was done by selection with collagenase (Fig. 1B)
Summary
A high efficacy of placental cells application necessitates their investigation. Preclinical studies require an improvement of the methods for obtaining, standardizing and storage of placental cells of experimental animals. Cells were isolated from rats and mice placentas by means of different enzymatic methods and the one of explants. The maximum number of viable cells from the placentas of mice and rats was found to be obtained using the explant method or trypsin with ETDA. The research results enabled the determining of effective biotechniques for obtaining the cryopreserved placental cells of rats and mice to perform preclinical studies of their biological effect in models of allo- and autotransplantations. Проте у клінічній практиці клітини плаценти людини передбачено використовувати в ало- та аутосистемах, що обумовлює необхідність проведення відповідних доклінічних досліджень та верифікації біобезпеки і потребує наявності ефективних біотехнологій отримання кріоконсервованих плацентарних клітин експериментальних тварин. У зв’язку з вищевикладеним метою даного дослідження були пошук і визначення ефективних методів виділення з плацент лабораторних тварин клітин для кріоконсервування, низькотемпературного зберігання і подальшого експериментального використання в моделях ало- та аутотрансплантації
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