Canãlulas-tronco de esperma humano cultivadas em laboratório, um passo inicial no tratamento da infertilidade
Uma idanãia para o tratamento da esterilidade masculina éa terapia com células-tronco espermatogonais (SSC).
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Ao aplicar um inibidor da molanãcula AKT, os pesquisadores da UC San Diego são capazes de favorecer a cultura de células-tronco espermatogonais humanas no laboratório, um primeiro passo em direção ao esperma produzido em laboratório como tratamento para a infertilidade masculina. Crédito: Vishaala Wilkinson
A infertilidade afeta um em cada sete homens em idade reprodutiva em todo o mundo. Uma idanãia para o tratamento da esterilidade masculina éa terapia com células-tronco espermatogonais (SSC). Nesta abordagem, as células-tronco dos espermatoza³ides no testaculo são transferidas para um tubo de ensaio, cultivadas e empurradas para se tornarem espermatoza³ides de pleno direito. No entanto, um gargalo importante foi identificar as condições corretas para que os SSCs humanos cresa§am no laboratório. Houve muitas tentativas, mas na maioria dos casos relatados não ficou claro se as células que estavam sendo cultivadas eram realmente SSCs, e nenhum manãtodo publicado anteriormente éusado rotineiramente.
Pesquisadores da Faculdade de Medicina da Universidade da Califórnia em San Diego agora desenvolveram um manãtodo confia¡vel para cultivar células com as caracteristicas dos SSCs humanos. Seu trabalho foi publicado na edição de 13 de julho de 2020 da Proceedings da Academia Nacional de Ciências .
"Achamos que nossa abordagem - que éapoiada por várias técnicas, incluindo a análise de sequenciamento de RNA de canãlula única - éum passo significativo para trazer a terapia com SSC para a clanica", disse o autor saªnior Miles Wilkinson, Ph.D., Distinguished Professor in Departamento de Obstetracia, Ginecologia e Ciências da Reprodução da Faculdade de Medicina da UC San Diego.
Os CSCs são o que tornam possível que os homens tenham filhos com mais de 65 anos. Essas células especializadas se auto-renovam continuamente, produzindo mais SSCs, e se desenvolvem em espermatoza³ides tão prolificamente que os homens (e algumas pessoas transgêneros, não bina¡rias e fluidas de gaªnero) produz mais de 1.000 novos espermatoza³ides a cada poucos segundos.
O progresso no campo foi dificultado pelo fato de ser extremamente difacil distinguir SSCs de outras células nos testaculos. Foi um grande passo em frente quando vários laboratórios, incluindo a equipe de Wilkinson, usaram recentemente uma técnica chamada seqa¼enciamento de RNA de canãlula única para definir as prova¡veis ​​caracteristicas moleculares especaficas dos CSCs humanos.
Em seu esfora§o mais recente, a equipe de Wilkinson usou suas informações de sequenciamento de RNA de canãlula única para purificar o que pensava serem SSCs humanos. Usando um manãtodo chamado transplante de células germinativas, ele mostrou que as células que purificavam eram de fato altamente enriquecidas em SSCs humanas. A equipe então reuniu o perfil dos genes expressos nesses SSCs humanos para fazer suposições sobre as condições que poderiam melhor apoiar seu crescimento no laboratório. Usando mais de 30 bia³psias de testaculo humano, os pesquisadores determinaram as condições certas necessa¡rias para cultivar células germinativas imaturas com as caracteristicas dos SSCs.
O principal ingrediente foi um inibidor da via AKT, um sistema celular que controla a divisão e a sobrevivaªncia celular. A equipe de Wilkinson determinou que a inibição do AKT mantanãm SSCs humanos, inibindo o desenvolvimento de precursores de espermatoza³ides em esta¡gio avana§ado . Atualmente, vários inibidores da AKT são usados ​​para tratar o ca¢ncer.
Com essa abordagem, os pesquisadores foram capazes de favorecer a cultura de células humanas com as caracteristicas moleculares dos SSCs por duas a quatro semanas.
"Em seguida, nosso principal objetivo éaprender a manter e expandir os CSCs humanos por mais tempo, para que possam ser clinicamente aºteis", disse Wilkinson.